Homeostatic capacity evaluation

ABSTRACT

Methods of evaluating homeostatic capacity of a subject are provided. Aspects of the methods include obtaining dynamic biometric data from the subject and evaluating the homoeostatic capacity of the subject from the obtained dynamic biometric data. Also provided are devices configured for use in practicing the methods. The methods and devices described herein find use in a variety of applications, e.g., health monitoring applications, treatment applications, dynamic diagnostic applications, etc.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application is a continuation-in-part application of U.S.application Ser. No. 15/061,645 filed on Mar. 4, 2016, whichapplication, pursuant to 35 U.S.C. § 119 (e), claims priority to U.S.Patent Provisional Patent Application Ser. No. 62/254,583 filed Nov. 12,2015; U.S. Provisional Patent Application No. 62/218,999 filed Sep. 15,2015; and U.S. Provisional Patent Application Ser. No. 62/128,816 filedMar. 5, 2015; the disclosures of which applications are hereinincorporated by reference.

INTRODUCTION

Homeostasis is a relatively stable state of equilibrium. Homeostaticcapacity (or allostatic capacity, buffering capacity, compensatorycapacity, or autoregulatory capacity) is the efficiency of anautoregulatory system to maintain functional homeostasis. Theinteraction of homeostatic capacity and stressors determines health.

Homeostatic capacity is so pervasively effective when we are young thatwe become more aware of it by its decline, especially after midlife,when recoveries from stressors—a late night, hangover, jet lag, cough,or injury—are readily noticeable. Our tolerance of contextual variation,such as changes in temperature and altitude, declines. Pupillary lightreflex response diminishes, and menus are harder to read in low light.Roller coasters and somersaults leave us more discombobulated as ourbiologic Weeble™ loses its self-centering efficiency.

While these are just a few of the subjective losses of homeostaticcapacity that we can feel, consider objective changes that we can'tfeel. The erosion of autoregulatory capacity of blood glucose, bloodpressure, tumor surveillance, and inflammation could partially explaindiabetes, hypertension, cancer, and various inflammatory diseases,respectively, independently of more widely accepted factors. Overall, ashomeostatic tolerance for internal derangements declines with age, thesame functions that promoted homeostasis during youth can behavedysfunctionally and promote secondary derangements in late adulthood(so-called antagonistic pleiotropy).

SUMMARY

Aspects of the present disclosure include a model for healthy,functional longevity based on evaluating and improving homeostaticcapacity. The inventors of the present application have realized thatvarious features of aging are epiphenomena of eroding homeostaticcapacity that reduce our ability to contend with ubiquitous stressorsand entropy. The inventors of the present application have realized thateroding homeostatic capacity is an Occam's razor that parsimoniouslyexplains the arborized taxonomy and diverse features of aging, and thatenhancing homeostatic capacity reverses the aging process and enablessustainable health.

Methods of evaluating homeostatic capacity of a subject are provided.Aspects of the methods include obtaining dynamic biometric data from thesubject and evaluating the homoeostatic capacity of the subject from theobtained dynamic biometric data. Also provided are devices configuredfor use in practicing the methods. The methods and devices describedherein find use in a variety of applications, e.g., health monitoringapplications, treatment applications, dynamic diagnostic applications,etc.

Aspects of the invention are based on the inventors' realization thatfor acute ailments, the proximate and ultimate goals of intervention aresymmetric—restoring homeostasis. During the pre-modern era, when humanstypically succumbed before senescence to acute insults such as traumaand infection, proximate interventions that acutely restored homeostasis(antibiotics, aseptic surgery) served patients well. Along with publichealth measures and vaccines, these treatments helped push mediansurvivorship past the age of female reproductive senescence. However,for chronic conditions that predominate today, including senescence,Whack-a-Mole™ acute remediations for chronic dysfunctions can induceatrophy of homeostatic capacity, leading to tachyphylaxis, nefariousfeed-forward decompensation, and progressive addiction to therapy. Manyeconomic stakeholders benefit from the current approach, but thelong-term benefit for patients is less clear. Extending lifespan withoutaddressing senescence creates an older population that consumes morehealth care, creating an unsustainable, feed-forward cycle of costescalation. A different framework is needed.

Aspects of the invention are based on the inventors' realization thatfor chronic conditions such as senescence (and others that occur atearlier ages), shifting the proximate goal to increasing homeostaticcapacity is a path to the ultimate restoration of functionalhomeostasis. This small conceptual change portends a radicaltransformation in the way we currently treat many chronic conditions.

For instance, hypertension today is typically medicated withantihypertensives, a method that seemingly resolves the problemproximately but is prone to induce tachyphylaxis ultimately. Embodimentsof the invention provide for administration of hormetic doses ofprohypertensive interventions that are pulsed to promote allostaticresponse—the way exercise does—to lower baseline blood pressure.

Embodiments of the invention include employing eustress (or capacitybuilding stress) to paradoxically promote homeostatic capacity as a wayto ultimately ameliorate various chronic diseases.

Aspects of the invention include the use of transformed lifestyleinterventions. Current static recommendations related to certain diets,exercise, stress reduction, and meditation are, in some instances,shifted to dynamic recommendations that increase the body's toleranceand dynamic range. With homeostatic capacity building as the new goal ofaspects of the present invention, lifestyle health emphasizes varianceover fixed habits. Meditation is oscillated with exhilaration. Exercisesvary in every dimension. Instead of eating a fixed diet such as vegan,some degree of variance that provides eustress is employed. Exposure tooscillations and variance in light and temperature, and other forms ofeustress, are encouraged.

Aspects of the invention further include fundamental resolutions—curesor near-cures where the benefits are sustained with reduced dependenceon repeat or ongoing intervention—for chronic conditions, includingsenescence. In that regard, embodiments of the invention employ editinggenes and pathways—e.g., as involved in biologic clocks, repairmechanisms, control systems, or energetics—that affect homeostaticcapacity. System dyssynchrony, loss of power, and desensitization tostimuli, which impair autoregulatory feedback loops and homeostaticcapacity, are among underlying common factors in cell aging that couldfractally connect cell senescence regulation to organismal senescence.

The present inventors have realized that dynamic systems such as biologyare better evaluated with dynamic models than static ones. Yet we haveheretofore been largely reliant on state variables as biomarkers. Annualchecks of heart rate, blood pressure, glucose, and cholesterol levels toassess health risks are tantamount to a seismic engineer relying on thedegree of angular declination, instead of resilience, to predictcollapse risk. Biologists, too, can add dynamic measures to currentstate variables to evaluate systems and homeostatic capacity—a conceptanalogous to resilience, strength, robustness, antifragility, andantifrailty. The present inventors have realized that biology is betterexplored four dimensionally, with more time series data and bettertemporal resolution. Aspects of the invention include a broad array ofdiagnostic tests with capabilities for continuous or high-frequencysampling.

In some instances, a system's potential homeostatic performance underall possible conditions, including every magnitude, type, pattern, andduration of stress, cannot be reduced into a single measure. However,approximations of homeostatic capacity are possible through a compositeof state variables, dynamic diagnostics, response diagnostics, andstress testing—assessment after contextual changes or allostatic loads.Examples of dynamic response tests include baroreceptor sensitivity,heart rate variability, flow-mediated vasodilation, postprandial glucoseand triglyceride tolerance tests, gait balance, pupillary light reflex,and recovery time after cardiac stress. In some embodiments, measures ofresilience correlate with chronologic age more than the underlying statevariables (example: heart rate recovery after cardiac stress versusresting heart rate).

Aspects of the invention include standards that allow approximations ofan individual's homeostatic capacity. Population-wide, age-basednormative values for a large panel of relatively uncorrelated measuresof homeostatic capacity may be employed. The normative values at peaklevels during youth can be used as benchmarks for understanding where anindividual's measures of homeostatic capacity stand as they advance inchronologic age. Alternatively, the measure of homeostatic capacity canbe personalized such that each individual's peak values can be used asself-referential benchmarks over time.

In some instances, homeostatic capacity levels beyond normative peaklevels are pursued not only by elite athletes and military personnel forthe purposes of human performance, but also by the broader populationfor the purposes of health, e.g., through training, capacity building,and intervention. Interventions that improve a large composite panel ofrelatively uncorrelated measures of homeostatic capacity—functionaloutcomes—are employed in some instances to obtain a systemic benefit.This category of interventions will fundamentally transform the futureof human health.

Homeostatic capacity is a fundamental trait of living systems, filteredby evolution at all scales of biology and all temporal epochs. Aspectsof the present disclosure include innovations that fundamentallyincrease homeostatic capacity which are a foundation to developingstrategies that will help avert the natural aging process and promotefunctional longevity. The end of aging would be the end of healthcare aswe know it. The feed-forward relationship between healthcare innovationand increasing future consumption would finally be decoupled.Restoration, augmentation and indefinite sustainment of homeostaticcapacity, e.g., as described herein, allows humans to persist at lowannual mortality rates currently enjoyed by young adults. Healthylifespan telescopes to a number of years that might have once seemedunimaginable. Human capacity is thus unleashed.

Aspects of the invention include employing a single intervention tomodulate, e.g., enhance, homeostatic capacity of two or more, such asthree or more, different, and in some instances seemingly unrelated,systems, e.g., as described above. For example, aspects of the inventioninclude employing a single intervention to improve the homeostaticcapacity of a composite of seemingly unrelated systems.

In some instances, homeostatic capacity is enhanced according to themethods described in U.S. application Ser. No. 15/363,980 entitled“Methods of Enhancing Homeostatic Capacity in a Subject by ModulatingHomeostatic System Synchrony, and Devices for Use in Practicing theSame,” the disclosure of which is herein incorporated by reference.

In some instances, homeostatic capacity is enhanced according to themethods described in U.S. application Ser. No. 15/363,988 entitled“Methods of Enhancing Homeostatic Capacity in a Subject by IncreasingHomeostatic System Component Responsiveness, and Devices for Use inPracticing the Same,” the disclosure of which is herein incorporated byreference.

BRIEF DESCRIPTION OF THE FIGURES

FIG. 1 is a flow chart illustrating one embodiment of a method forevaluating homeostatic capacity of a subject.

FIG. 2 is a flow chart illustrating one embodiment of a method forevaluating homeostatic capacity of a subject, specifically showing amachine learning algorithm used in classification.

DETAILED DESCRIPTION

Methods of evaluating homeostatic capacity of a subject are provided.Aspects of the methods include obtaining dynamic biometric data from thesubject and evaluating the homoeostatic capacity of the subject from theobtained dynamic biometric data. Also provided are devices configuredfor use in practicing the methods. The methods and devices describedherein find use in a variety of applications, e.g., health monitoringapplications, treatment applications, dynamic diagnostic applications,etc.

Before the present invention is further described, it is to beunderstood that this invention is not limited to particular embodimentsdescribed, as such may, of course, vary. It is also to be understoodthat the terminology used herein is for the purpose of describingparticular embodiments only, and is not intended to be limiting, sincethe scope of the present invention will be limited only by the appendedclaims.

Where a range of values is provided, it is understood that eachintervening value, to the tenth of the unit of the lower limit unlessthe context clearly dictates otherwise, between the upper and lowerlimit of that range and any other stated or intervening value in thatstated range, is encompassed within the invention. The upper and lowerlimits of these smaller ranges may independently be included in thesmaller ranges and are also encompassed within the invention, subject toany specifically excluded limit in the stated range. Where the statedrange includes one or both of the limits, ranges excluding either orboth of those included limits are also included in the invention.

Methods recited herein may be carried out in any order of the recitedevents which is logically possible, as well as the recited order ofevents.

Unless defined otherwise, all technical and scientific terms used hereinhave the same meaning as commonly understood by one of ordinary skill inthe art to which this invention belongs. Although any methods andmaterials similar or equivalent to those described herein can also beused in the practice or testing of the present invention, the preferredmethods and materials are now described.

It must be noted that as used herein and in the appended claims, thesingular forms “a”, “an”, and “the” include plural referents unless thecontext clearly dictates otherwise. It is further noted that the claimsmay be drafted to exclude any optional element. As such, this statementis intended to serve as antecedent basis for use of such exclusiveterminology as “solely,” “only” and the like in connection with therecitation of claim elements, or use of a “negative” limitation.

All publications mentioned herein are incorporated herein by referenceto disclose and describe the methods and/or materials in connection withwhich the publications are cited.

The publications discussed herein are provided solely for theirdisclosure prior to the filing date of the present application. Nothingherein is to be construed as an admission that the present invention isnot entitled to antedate such publication by virtue of prior invention.Further, the dates of publication provided may be different from theactual publication dates which may need to be independently confirmed.

In further describing the invention, aspects of embodiments of methodsof evaluating homeostatic capacity are described first in greaterdetail, followed by a description of representative devices that finduse in practicing such methods. Next, a review of various applicationsin which the methods and devices find use, is provided.

Methods of Homeostatic Capacity Evaluation

As summarized above, methods for evaluating homeostatic capacity of asubject are provided. Homeostatic capacity refers to the ability of asubject to maintain relatively constant conditions in the internalenvironment while continuously interacting with and adjusting to changesoriginating within or outside the system. By “evaluating” is meantassessing, analyzing or assaying to provide a form of measurement, e.g.,in the form of a determination or proxy thereof, of the homeostaticcapacity of the subject. The evaluations that may be made may bequantitative and/or qualitative determinations, and be represented as avalue or set of values, as desired.

Aspects of the methods include obtaining dynamic biometric data from asubject. The phrase “biometric data” is employed to refer to a measureof a biometric parameter that relates to the physiology of a livingorganism, e.g., as described below. As such, the biometric parameterwhich is employed in methods of the invention to obtain the biometricdata may be a parameter that provides information about an organism'svital functions, including growth and development, the absorption andprocessing of nutrients, the synthesis and distribution of proteins andother organic molecules, the functioning of different tissues, organs,and other anatomic structures; the psychological and/or behavioral stateof the subject, e.g., mental and/or cognitive state of the subject,which may be subjective or objective, self-reported or third partyobserved, as desired; etc.

Biometric parameters that are measured may vary widely, where examplesof such parameters include physiological, chemical, electrical,behavioral, psychological, etc., based parameters, as well as variationsand derivatives thereof. Biometric parameters of interest include, butare not limited to: physical parameters, e.g., blood pressure,orthostatic hypotension, pulse pressure, heart rate, heart ratevariability (HRV), heart rate recovery, resting heart rate, respirationrate, forced expiratory volume, forced vital capacity, temperature, coretemperature, galvanic skin response, gastrointestinal motility, sleepcycle, VO2 max, bone density, weight, body mass index (BMI), bonedensity, waist to hip ratio, waist circumference, other obesity measures(e.g., volume displacement, Dual Energy X-ray Absorptiometry (i.e.,DEXA), etc.), baroreceptor sensitivity, oxygen saturation, nervoussystem activity measurements, including electrical potentialmeasurements, such as spontaneous electrical potential measurements,e.g., EEG, EMG EKG, evoked electrical potential measurements, e.g.,sensory evoked potentials (such as auditory invoked potentials (e.g.,brain stem evoked response or potential (ABER or ABEP), visual evokedpotentials, tactile or somatosensory evoked potentials, laser evokedpotentials), motor evoked potentials, etc.; nerve conduction measures,e.g., motor NCS, sensory NCS, F-wave study, H-reflex study, spf-NCS,etc.; and the like, and combinations thereof; sample analysis obtainableparameters, e.g., pH level, cortisol level, ACTH level,Epinephrine/Norepinephrine level, oxygen saturation, insulin, glucose,inflammatory/immune markers, DNA methylation, DNA double strand breaks,clock genes/factors, oxidative stress, telomere status, gut biome,melatonin level, adenosine level, creatinine, urea nitrogen, c-reactiveprotein, hemoglobin, triglycerides, lipoproteins, apolipoprotein B100/A1ratio, white blood cell count, cholesterol, oxygen saturation, and thelike, and combinations thereof. Dynamic biometric data may be made up ofinformation about a single type of biometric parameter, or two or moredifferent types of biometric parameters. The biometric data employed inmethods of the invention may thus be made up of information obtained bymeasuring or assessing one or more biometric parameters, such as theones listed above.

As summarized above, the biometric data that is obtained and employed inmethods of the invention is dynamic biometric data. By “dynamicbiometric data” is meant biometric data that incorporates some type ofchange component, as opposed to static biometric data. The changecomponent may vary widely, where examples of change components include,but are not limited to components that are: temporal and/or in responseto an applied stimulus and/or in response to withdrawal of stimulusand/or in response to a change in the contextual environment of thesubject. For example, the dynamic biometric data that is obtained may bebiometric data obtained over a given period of time. The given period oftime may vary, ranging in some instances from 0.1 seconds to 24 hours,such as 1 second to 12 hours, e.g., 1 second to 1 hour, including 1second to 1 minute. Where the dynamic biometric data is data obtainedover a given period of time, the data may be obtained continuously overthat period of time or at one or more distinct points during that periodof time. For example, the biometric parameter(s) that is monitored inorder to obtain biometric data may be monitored continuously during thegiven period of time, i.e., it may be obtained in an uninterruptedmanner, i.e., without cessation, during the given period of time.Alternatively, the biometric parameter(s) that is monitored in order toobtain biometric data may be monitored intermittently during the givenperiod of time, i.e., it may be obtained at one or more points over thegiven period of time, with an interval between points at which it is notobtained. In some embodiments, the interval may vary, ranging, forexample, from 0.01 sec to 60 minutes or longer, such as 0.1 to 60 s. Insome instances, the dynamic biometric data is obtained by evaluating abiometric parameter for a rate of change over a period of time. As such,methods may include obtaining information about the speed at which abiometric parameter of interest changes over a given period of period oftime. Obtaining dynamic biometric data as described above provides fornumerous benefits, including increases in temporal resolution, ascompared to single point in time data. Dynamic biometric data asobtained herein provides a truer and more meaningful measure of thebiometric value(s) of interest, as compared to single point in timemeasurements.

Dynamic biometric data of interest also includes biometric data that isobtained by evaluating a biometric parameter for a change in response toan applied stimulus. Such biometric data may include data that isobtained before and/or after application of the stimulus to the subject.In some instances, the biometric data may be obtained over a givenperiod of time that spans or follows the application of the stimulus tothe subject. This type of biometric data may be viewed as biometric datathat is obtained over a given period of time in conjunction withapplication of a stimulus to the subject being evaluated. The appliedstimulus may vary, where stimuli of interest include physical stimuliand chemical stimuli. Physical stimuli of interest include, but are notlimited to, change in orientation of the subject, exercise, change intemperature experienced by the subject or a portion thereof, and thelike. Chemical stimuli of interest include, but are not limited to,administration of various active agents, e.g., orally, topically, byinjection or other type of administration route, where active agents ofinterest include, but are not limited to: sugars, starches, stimulants,and the like.

Dynamic biometric data of interest also includes biometric data that isobtained by evaluating a biometric parameter for a change in response towithdrawal of a stimulus. Such biometric data may include data that isobtained before and/or after withdrawal (e.g., blockage) of the stimulusto the subject. In some instances, the biometric data may be obtainedover a given period of time that spans or follows the withdrawal of thestimulus to the subject. This type of biometric data may be viewed asbiometric data that is obtained over a given period of time inconjunction with withdrawal of a stimulus to the subject beingevaluated. The withdrawn stimulus may vary, where stimuli of interestinclude physical stimuli and chemical stimuli. Physical stimuli ofinterest include, but are not limited to, change in orientation of thesubject, exercise, change in temperature experienced by the subject or aportion thereof, and the like. Chemical stimuli of interest include, butare not limited to, administration of various active agents, e.g.,orally, topically, by injection or other type of administration route,where active agents of interest include, but are not limited to: sugars,starches, stimulants, and the like.

Dynamic biometric data of interest also includes biometric data that isobtained by evaluating a biometric parameter for a change in response tomodulation of the contextual environment of the subject. By contextualenvironment of the subject is meant the perceived environment of thesubject. Such biometric data may include data that is obtained beforeand/or after the modulation in the contextual environment of thesubject. In some instances, the biometric data may be obtained over agiven period of time that spans or follows the modulation of thecontextual environment of the subject. This type of biometric data maybe viewed as biometric data that is obtained over a given period of timein conjunction with modulation of the contextual environment of thesubject. The modulation of the contextual environment of the subject mayvary, where contextual modulations of interest include, but are notlimited to, change in day and night duration, change in temperature,change in humidity, change in elevation, change in atmosphere, and thelike.

Dynamic biometric data of interest also includes biometric data that isobtained by evaluating a biometric parameter for a change in response tomodulation of the behavioral aspect of the subject. By behavioral aspectof the subject is meant an observable activity of the subject. Suchbiometric data may include data that is obtained before and/or after themodulation of the behavioral aspect of the subject. In some instances,the biometric data may be obtained over a given period of time thatspans or follows the modulation of the behavioral aspect of the subject.This type of biometric data may be viewed as biometric data that isobtained over a given period of time in conjunction with modulation ofthe behavioral aspect of the subject. The modulation of the behavioralaspect of the subject may vary, where behavioral modulations of interestinclude, but are not limited to, dietary changes, sleep pattern changes,activity level changes, and the like. It is noted that the dynamicbiometric data that is employed in methods of the invention may be thedata that is directly obtained from a suitable sensor, or a derivativethereof. As such, derivative measurements of dynamic biometric data maybe obtained in methods of the invention, include second, third, etc.order derivative data. For example, instead of or in addition toemploying a measured change in a parameter, such as acceleration ordeceleration, one may employ the speed of the measured change in theparameter, i.e. first derivative.

As reviewed above, a variety of different biometric parameters may bemeasured to obtain the dynamic biometric data. The method by which thebiometric data is obtained may vary depending on the nature of thebiometric parameter that is monitored. In some instances, the methodemployed to obtain the biometric data includes physically monitoring thesubject to obtain the dynamic biometric data. For example, physicalmonitoring of the subject may be employed where the biometric parameteris one or more of blood pressure, orthostatic hypotension, pulsepressure, heart rate, heart rate variability (HRV), heart rate recovery,resting heart rate, respiration rate, forced expiratory volume, forcedvital capacity, temperature, core temperature, galvanic skin response,gastrointestinal motility, sleep cycle, VO2 max, bone density, weight,body mass index (BMI), bone density, waist to hip ratio, waistcircumference, other obesity measures (e.g., volume displacement, DualEnergy X-ray Absorptiometry (i.e., DEXA), etc.), baroreceptorsensitivity, oxygen saturation, nervous system activity measurements,including electrical potential measurements, such as spontaneouselectrical potential measurements, e.g., EEG, EMG EKG, evoked electricalpotential measurements, e.g., sensory evoked potentials (such asauditory invoked potentials (e.g., brain stem evoked response orpotential (ABER or ABEP), visual evoked potentials, tactile orsomatosensory evoked potentials, laser evoked potentials), motor evokedpotentials, etc.; nerve conduction measures, e.g., motor NCS, sensoryNCS, F-wave study, H-reflex study, spf-NCS, etc.; and combinationsthereof. Any convenient protocol for physically monitoring a subject forone or more of the above biometric parameters may be employed, andmethods for physically monitoring each are known in the art. Forexample, where the biometric parameter of interest is HRV, the physicalmonitoring may include measures such as low frequency peak (“LF”), highfrequency peak (“HF”), and the LF/HF ratio to determine HRV and obtainthe HRV derived biometric data.

In some embodiments, the dynamic biometric data is obtained by a methodthat includes analyzing a sample from the subject to obtain the dynamicbiometric data. The sample that is analyzed may vary, where samples ofinterest include, but are not limited to: urine, blood, serum, plasma,saliva, semen, prostatic fluid, nipple aspirate fluid, lachrymal fluid,perspiration, feces, cheek swabs, cerebrospinal fluid, cell lysatesamples, amniotic fluid, gastrointestinal fluid, biopsy tissue (e.g.,samples obtained from laser capture microdissection (LCM)), and thelike. The sample can be a biological sample or can be extracted from abiological sample derived from humans, animals, and the like, and mayemploy conventional methods for the successful extraction of DNA, RNA,proteins and peptides. In certain embodiments, the sample is a fluidsample, such as a solution of analytes in a fluid. The fluid may be anaqueous fluid, such as, but not limited to water, a buffer, and thelike. Biometric parameters that may be monitored by evaluating a samplefrom the subject include, but are not limited to: pH level, cortisollevel, ACTH level, Epinephrine/Norepinephrine level, oxygen saturation,insulin, glucose, inflammatory/immune markers, DNA methylation, DNAdouble strand breaks, clock genes/factors, oxidative stress, telomerestatus, gut biome, melatonin level, adenosine level, creatinine, ureanitrogen, c-reactive protein, hemoglobin, triglycerides, lipoproteins,apolipoprotein B100/A1 ratio, white blood cell count, cholesterol,oxygen saturation, and combinations thereof.

Any convenient protocol for physically monitoring a subject for one ormore of the above biometric parameters may be employed, and methods fortesting a sample for monitoring each are known in the art. In someinstances, the dynamic biometric data is obtained by both physicallymonitoring the subject and by assaying a sample from the subject, e.g.,as described above. Of interest in certain embodiments are virtualreality (VR) mediated protocols.

Aspects of the methods further include evaluating the homoeostaticcapacity of the subject from the dynamic biometric data. As such,following obtainment of the dynamic biometric data, the homeostaticcapacity of the subject is evaluated based on the obtained dynamicbiometric data. Any convenient protocol may be employed to evaluate thehomeostatic capacity of the subject based on the obtained dynamicbiometric data. For example, the obtained dynamic biometric data may becompared to control or reference sets of dynamic biometric data toobtain the homeostatic capacity evaluation. In some instances, theobtained dynamic biometric data may be compared to a suitable databaseof control or reference sets to obtain the homeostatic capacityevaluation. The control or references sets of data may be made up ofdata obtained from multiple different individuals of known homeostaticcapacity. The data may be made up from individuals of a variety ofdifferent ages and health, including from young and old individuals, aswell as healthy and diseased individuals, as desired. Any suitablecomparison algorithm may be employed, and the output homeostaticcapacity evaluation may be produced in a variety of different formats orconfigurations. This homeostatic capacity evaluation step may beperformed using a suitable functional module of a computingdevice/system, e.g., as described in greater detail below.

The homeostatic capacity evaluation that is provided by methods of theinvention may vary, as desired. For example, the evaluation may be anoutput in the form of a qualitative assessment, e.g., bad, poor,average, good and exceptional, etc. The output may be in the form of aquantitative assessment, e.g., where the homeostatic capacity evaluationoutput a number selected from a numerical scale. The homeostaticcapacity evaluation output may provide assessment with respect to anumber of different homeostatic capacity parameters, such as but notlimited to: the robustness, dynamic range, resilience, coping mechanism,anti-fragility, etc., of the homeostatic capacity of the individual. Theoutput showing the homeostatic capacity of the animal/person may beprovided as a proxy for the biological age (as opposed to thechronological age) of the subject, e.g., by using statisticalcorrelations relative to the general population. For example, thehomeostatic capacity evaluation produced from dynamic biometric datafrom a 50 year old professional cyclist in great condition could suggestthat the “biological age” of that person based on homeostatic capacitymeasures is actually much younger, e.g., that of a 35 years old from thegeneral population. In some instances, the homeostatic capacityevaluation is one that is prepared by comparing the obtained dynamicbiometric data to a database that includes data comprising statisticallymeaningful values that correlate each biometric value and/or acombination of the biometric values of interest to the values ofdifferent ages or age ranges of cohorts for the same biometric value(s).For example, in instances where the obtained biometric data may be froman individual or animal that is 30 years of age, the homeostaticcapacity evaluation may be performed by comparing the obtained biometricdata to data obtained from healthy individuals from a variety of agesranging from 20 to 80 years, and show a correlation to a certain age ofthe individual as a whole or certain systems thereof, e.g.,cardiovascular system, neurological system, reproductive system, etc.For example, the output homeostatic capacity evaluating may be anoverall composite number, e.g., that the individual has the homeostaticcapacity of a 32 year old, or be more granular with respect toparticular biological systems of the individual, e.g., where the outputis that the system provides a homeostatic capacity evaluation in whichthe subject has a cardiovascular system of a 25 year old but the nervoussystem of a 35 year old. In such instances, these sub-categories couldbe at systems levels of the body and could be more granular, e.g.,portions of systems.

The methods described herein may be employed with a variety of differenttypes of subjects, i.e., animals, where the animals may be “mammals” or“mammalian,” where these terms are used broadly to describe organismswhich are within the class mammalia, including the orders carnivore(e.g., dogs and cats), rodentia (e.g., mice, guinea pigs, and rats),lagomorpha (e.g., rabbits) and primates (e.g., humans, chimpanzees, andmonkeys). In some instances, the subjects or patients are humans orlaboratory research animals.

In some embodiments, the methods may further include modulating thehomoeostatic capacity of the subject following obtainment of thehomeostatic capacity evaluation of the subject. For example, the methodsmay include at least partially restoring the homeostatic capacityfollowing evaluation of such. By “at least partially restoring thehomeostatic capacity of the subject” is meant that the homeostaticcapacity of the subject is enhanced or improved, e.g., to that of atarget value, which target value may be a “normal” value or greater thana normal value, e.g., a super-normal value. By “normal” is meant thehomeostatic capacity of a healthy subject of a particular age. Incertain embodiments, the healthy subject is a healthy human at an ageafter puberty, e.g., 18 year old, 19 year old, 20 year old, 21 year old,22 year old, 23 year old, 24 year old, 25 year old, 26 year old, 27 yearold, 28 year old, 29 year old, 30 year old, 31 year old, 32 year old, 33year old 34 year old, 35 year old, 36 year old, 37 year old, 38 yearold, 39 year old, 40 year old, 41 year old, 42 year old, 43 year old, 44year old, 45 year old, 46 year old, 47 year old, 48 year old, 49 yearold or 50 year old. In some instances, the normal function with respectto homeostatic capacity is that of a healthy human 25 year old. By supernormal value is meant the homeostatic capacity of a subject of havegreater than normal homeostatic capacity, e.g., that of an athlete, etc.The magnitude of difference between normal and super normal may vary,and in some instances may be 5% or greater, such as 10% or greater,including 15%, 20% or 25% or greater, where in some instances the targetsuper normal homeostatic capacity is 5% to 75% greater than of a normalhomeostatic capacity. In some instances, the methods include modulatingthe homeostatic capacity of the subject to that it is at least closer toa target homeostatic capacity. By “at least closer” is meant, in someinstances, that the target homeostatic capacity is restored to be 50% ormore, e.g., 75% or more of the target function, such as 80% or more ofthe target function, including 90% or more of the target function, e.g.,95% or more of the target function, including 99% or more of the targetfunction.

Modulation of the homeostatic capacity of a subject as described abovecan be achieved using any suitable protocol, including, but not limitedto electrical and/or pharmacologic and/or physical and/or chemicaland/or psychological and/or environmental and/or behavioral protocols,e.g., as described in greater detail below.

In some instances, the methods may include use of one or more staticmeasures of homeostatic capacity. Such measures may be used as separatemeasures, or composites of dynamic and static measurements may beemployed.

Embodiments of the methods result in rapid restoration of homeostaticcapacity of the subject. For example, in some instances homeostaticcapacity may be restored in 72 hours or less relative to the onset ofautonomic modulation, such as in 48 hours or less, 24 hours or less, or12 hours or less relative to the onset of autonomic modulation. In otherembodiments of the methods, the restoration of homeostatic capacity maytake a longer period of time, e.g., 1 or more days or longer, 1 or moremonths or longer, including 1 or more years or longer. The therapeuticprotocol employed may be continuously or periodically applied, e.g.,where the protocol is mediated by an implanted device that provides forcontinuous, intermittent or period administration of electrical and/orpharmacological treatment, e.g., as described in greater detail below.

Utility

The subject methods find use in a variety of different applications.

Applications of interest include, but are not limited to: health andwellness monitoring applications; diagnostic applications; preventativeapplications; treatment applications; etc.

Health/Wellness Applications

For example, the methods described herein may be employed in varioushealth and wellness monitoring applications, e.g., by individualsmonitoring themselves or interested stakeholders, e.g., health careprofessionals, physical trainers, family or friends, etc., monitoringthe individuals. For example, the methods may be employed by individualsto monitor their homeostatic capacity on an ongoing basis, e.g., so thatthey can monitor their health and well-being over time. The individualsmay use the homeostatic capacity evaluation to make lifestyle changes,e.g., changes in diet and/or exercise. Alternatively or in addition, themethods may be employed by a stakeholder having an interest in thehealth of an individual, such as the stakeholders listed above.

Diagnostic Applications

In yet other instances, the methods may be employed in at least theprediction of the presence of, if not diagnosis of, a condition that ispresent or may occur in a subject, which may be a disease or othercondition, such as the conditions described below. For example, in someembodiments the homeostatic capacity evaluation or underlying datathereof, e.g., dynamic measurement data, may be employed to predict, ifnot diagnose, the presence of a disease condition in the subject. Insome instances, the methods may be employed to predict thepredisposition of a subject for developing a disease condition in thefuture, and may further include developing a regimen configured to delayor prevent occurrence of the predicted condition. In some instances, themethods may be employed to predict the impending occurrence in a subjecta disease condition or symptom, e.g., episode or acute condition,thereof, e.g., of an allergic, e.g., anaphylaxis, episode; an autisticepisode; etc. In such dynamic diagnostic applications, any convenientdynamic measure may be employed, such as those described above, e.g.,heart rate variability readings, EEG readings, EKG readings, etc. Thedynamic measure data may be employed raw in an algorithm used to predictthe impending occurrence, or may be processed and then used to predictthe impending occurrence, e.g., an HC evaluation obtained from the rawdata may be employed to predict the impending occurrence, as desired.For example, dynamic diagnostic applications of the invention includeobtaining dynamic measures of one or more of HRV, EEG and EKG, e.g., asdescribed above, and using the resultant data to predict the impendingoccurrence of a condition symptom or episode, e.g., an allergicresponse, e.g., an anaphylactic response, where the prediction may bemade some time prior to the occurrence, e.g., 5 seconds or more, 10seconds or more, 30 seconds or more, 60 seconds or more, 5 minutes ormore, 10 minutes or more, 15 minutes or more, 30 minutes or more, 60minutes or more, 2 hours or more, 6 hours or more, 12 hours or more, 24hours or more, before the occurrence of the condition symptom orepisode. In such instances, the prediction of the future occurrence maybe coupled with appropriate therapeutic intervention, e.g., to preventthe occurrence, ameliorate the magnitude of the occurrence, reduce theimpact on others of the occurrence, etc.

Treatment Applications

In some instances, the methods find use as a component of the treatmentof a variety of different conditions. By treatment is meant that atleast an amelioration of the symptoms associated with the conditionafflicting the subject is achieved, where amelioration is used in abroad sense to refer to at least a reduction in the magnitude of aparameter, e.g. symptom, associated with the condition being treated. Assuch, treatment also includes situations where the condition, or atleast symptoms associated therewith, are completely inhibited, e.g.prevented from happening, or stopped, e.g. terminated, such that thesubject no longer suffers from the condition, or at least the symptomsthat characterize the condition. In certain embodiments, the conditionbeing treated is a disease condition.

Non-limiting examples of disease conditions that may be treated bypractice of the methods include, but are not limited to: Examples ofconditions that may be treated with the methods of the subject inventioninclude, but are not limited to, cardiovascular diseases, e.g.,atherosclerosis, coronary artery disease, hypertension, hyperlipidemia,cardiomyopathy, volume retention; neurodegenerative diseases, e.g.,Alzheimer's disease, Pick's disease, dementia, delirium, Parkinson'sdisease, amyotrophic lateral sclerosis; neuroinflammatory diseases,e.g., viral meningitis, viral encephalitis, fungal meningitis, fungalencephalitis, multiple sclerosis, charcot joint; myasthenia gravis;orthopedic diseases, e.g., osteoarthritis, inflammatory arthritis,reflex sympathetic dystrophy, Paget's disease, osteoporosis;lymphoproliferative diseases, e.g., lymphoma, lymphoproliferativedisease, Hodgkin's disease; autoimmune diseases, e.g., Graves disease,hashimoto's, takayasu's disease, kawasaki's diseases, arthritis,scleroderma, CREST syndrome, allergies, dermatitis, Henoch-schlonleinpurpura, goodpasture syndrome, autoimmune thyroiditis, myastheniagravis, Reiter's disease, lupus, rheumatoid arthritis; inflammatory andinfectious diseases, e.g., sepsis, viral and fungal infections, woundhealing, tuberculosis, infection, human immunodeficiency virus;pulmonary diseases, e.g., tachypnea, fibrotic diseases such as cysticfibrosis, interstitial lung disease, desquamative interstitialpneumonitis, non-specific interstitial pneumonitis, lymphocyticinterstitial pneumonitis, usual interstitial pneumonitis, idiopathicpulmonary fibrosis; transplant-related side effects such as rejection,transplant-related tachycardia, renal failure, typhlitis; transplantrelated bowel dysmotility, transplant-related hyperreninemia; sleepdisorders, e.g., insomnia, obstructive sleep apnea, central sleep apnea;gastrointestinal disorders, e.g., hepatitis, xerostomia, boweldysmotility, peptic ulcer disease, constipation, post-operative boweldysmotility; inflammatory bowel disease; endocrine disorders, e.g.,hypothyroidism, hyperglycemia, diabetes, obesity, syndrome X; cardiacrhythm disorders, e.g., sick sinus syndrome, bradycardia, tachycardia,QT interval prolongation arrhythmias, atrial arrhythmias, ventriculararrhythmias; genitourinary disorders, e.g., bladder dysfunction, renalfailure, hyperreninemia, hepatorenal syndrome, renal tubular acidosis,erectile dysfunction; cancer; fibrosis; skin disorders, e.g., wrinkles,cutaneous vasculitis, psoriasis; aging associated diseases andconditions, e.g., shy dragers, multi-system atrophy, osteoporosis, agerelated inflammation conditions, degenerative disorders; autonomicdysregulation diseases; e.g., headaches, concussions, post-concussivesyndrome, coronary syndromes, coronary vasospasm; neurocardiogenicsyncope; neurologic diseases such as epilepsy, seizures, stress, bipolardisorder, migraines and chronic headaches; conditions related topregnancy such as amniotic fluid embolism, pregnancy-relatedarrhythmias, fetal stress, fetal hypoxia, eclampsia, preeclampsia;conditions that cause hypoxia, hypercarbia, hypercapnia, acidosis,acidemia, such as chronic obstructive lung disease, emphysema,cardiogenic pulmonary edema, non-cardiogenic pulmonary edema, neurogenicedema, pleural effusion, adult respiratory distress syndrome,pulmonary-renal syndromes, interstitial lung diseases, pulmonaryfibrosis, and any other chronic lung disease; sudden death syndromes,e.g., sudden infant death syndrome, sudden adult death syndrome;vascular disorders, e.g., acute pulmonary embolism, chronic pulmonaryembolism, deep venous thrombosis, venous thrombosis, arterialthrombosis, coagulopathy, aortic dissection, aortic aneurysm, arterialaneurysm, myocardial infarction, coronary vasospasm, cerebral vasospasm,mesenteric ischemia, arterial vasospasm, malignant hypertension; primaryand secondary pulmonary hypertension, reperfusion syndrome, ischemia,cerebral vascular accident, cerebral vascular accident and transientischemic attacks; pediatric diseases such as respiratory distresssyndrome; bronchopulmonary dysplasia; Hirschprung disease; congenitalmegacolon, aganglionosis; ocular diseases such as glaucoma; and thelike.

In some instances, at least partial restoration of the homoeostaticcapacity of a subject results in treatment of a condition caused bysympathetic bias. Conditions that are caused by a sympathetic biasinclude, but are not limited to aging related diseases, such as but notlimited to: cardiovascular disease, cancer, arthritis, cataracts,osteoporosis, type 2 diabetes, hypertension; shy dragers, multi-systematrophy, age related inflammation conditions and diabetes.

In some instances, at least partial restoration of the homoeostaticcapacity of a subject results in treatment of a condition caused byparasympathetic bias. Conditions that are caused by a parasympatheticbias include, but are not limited to an allergy, common cold, eczema,asthma, anaphylaxis, attention deficit hyperactive disorder (ADHD),autism, obesity, depression, Tourette's syndrome, hay fever, cough,fatigue, hypothyroidism, chronic fatigue syndrome, environmentalsensitivity syndrome, shock, sepsis, food allergy and food allergysyndrome.

Also of interest are non-disease conditions, where such non-diseaseconditions include, but are not limited to: aging, sleep deprivation,veisalgia, and the like.

As such, aspects of the invention include methods that further includetreating a subject for a condition. Embodiments of such methods include:obtaining a dynamic measure of homeostatic capacity for the subject,e.g., as described above, and administering a therapy to the subject ina manner sufficient to modulate the subject's dynamic measure ofhomeostatic capacity to more closely approximate a target dynamicmeasure of homeostatic capacity and treat the subject for the condition.In some instances, the methods may include a homeostatic capacitymeasurement that is based on one or more static measures of homeostaticcapacity. Such measures may be used as separate measures, or compositesof dynamic and static measurements may be employed.

In these embodiments of the invention, any convenient therapy may beadministered to a subject. Therapies that may be employed include, butare not limited to: traditional medical therapies, e.g., electricaltherapies, pharmacological therapies, electropharmaceuticals, etc.; andnon-traditional medical therapies, e.g., homeopathic therapies,acupuncture, acupressure, mechanical manipulation, e.g., chiropractictherapies, laser therapy, e.g., to the vertex or other physiologicallocations, etc. Therapies of interest may also be categorized asphysical, chemical, psychological, environmental, electrical,behavioral, pharmacological, etc. Specific types of therapies ofinterest are now reviewed in greater detail.

ANS Modulation

In some instances, the administered therapy is one that modulates theautonomic nervous system of the subject. The autonomic nervous system(“ANS”) is that portion of the nervous system that is not the somaticnervous system. The ANS controls individual organ function andhomeostasis. For the most part, the ANS is not subject to voluntarycontrol. The ANS is also commonly referred to as the visceral orautomatic system. The ANS can be viewed as a “real-time” regulator ofphysiological functions that extracts features from the environment and,based on that information, allocates an organism's internal resources toperform physiological functions for the benefit of the organism, e.g.,responds to environment conditions in a manner that is advantageous tothe organism. The ANS conveys sensory impulses to and from the centralnervous system to various structures of the body such as organs andblood vessels, in addition to conveying sensory impulses through reflexarcs. For example, the ANS controls constriction and dilatation of bloodvessels; heart rate; the force of contraction of the heart; contractionand relaxation of smooth muscle in various organs; lungs; stomach;colon; bladder; visual accommodation, secretions from exocrine andendocrine glands, etc. The ANS does this through a series of nervefibers and more specifically through efferent and afferent nerves.

The ANS acts through a balance of its two components: the sympatheticnervous system and parasympathetic nervous system, which are twoanatomically and functionally distinct systems. Both of these systemsinclude myelinated preganglionic fibers which make synaptic connectionswith unmyelinated postganglionic fibers, and it is these fibers whichthen innervate the effector structure. These synapses usually occur inclusters called ganglia. Most organs are innervated by fibers from bothdivisions of the ANS, and the influence is usually opposing (e.g., thevagus nerve slows the heart, while the sympathetic nerves increase itsrate and contractility), although it may be parallel (e.g., as in thecase of the salivary glands).

By “modulating” is meant altering or changing one or more aspects orcomponents to provide a change, alteration or shift in another aspect orcomponent. Modulating autonomic function is achieved by modulating atleast one portion of the subject's autonomic nervous system. By“modulating at least one portion of the subject's autonomic nervoussystem” is meant altering or changing at least a portion of an autonomicnervous system by a means to provide a change, alteration or shift in atleast one component or aspect of the autonomic nervous system.

In some instances of the subject methods, modulation of the autonomicnervous system includes modulating the parasympathetic and/orsympathetic activity in the subject. “Parasympathetic activity” refersto activity of the parasympathetic nervous system whereas “sympatheticactivity” refers to activity of the sympathetic nervous system.

In some instances, modulation results in at least one of decreasingparasympathetic activity and/or increasing sympathetic activity in asubject to improve a condition caused by parasympathetic bias. In otherembodiments, the modulation results in at least one of decreasingsympathetic activity and/or increasing parasympathetic activity in asubject to improve a condition caused by sympathetic bias.

Therapeutic modalities may employ modulation of activity in or morecomponents of the nervous system. The nervous system includes the spinalcord and the pairs of nerves along the spinal cord which are known asspinal nerves. The spinal nerves include both dorsal and ventralbranches which fuse in the intravertebral foramen to create a mixednerve. Methods employed in the invention may modulate only one of thedorsal or ventral branches, or both of the dorsal and ventral branches,where when both of the dorsal and ventral branches are modulated, themodulation may be the same or different, e.g., where the two branchesare differentially modulated.

Modulation of the autonomic nervous system may be carried out using anysuitable protocol, including, but not limited to: electrical and/orpharmacologic and/or physical and/or chemical and/or psychologicaland/or environmental protocols, e.g., as described below. The modulationof the ANS provides, in some instances, an increase in function of atleast a portion of the autonomic system, e.g., increase function in atleast one sympathetic or parasympathetic nerve fiber, and/or provides,in some instances, a decrease in function or dampening of a portion ofthe autonomic system, e.g., may inhibit activity in at least onesympathetic or parasympathetic nerve fiber or inhibit nerve pulsetransmission.

In some instances, the modulation that is achieved in practicing methodsof the invention may be quantified. One way of quantifying modulation ofat least one portion of the subject's autonomic nervous system is theparasympathetic/sympathetic activity ratio. By“parasympathetic/sympathetic activity ratio” is meant the ratio ofactivity of the sympathetic nervous system to the activity of theparasympathetic nervous system. As such, methods according to certainembodiments include modulating a sympathetic/parasympathetic activityratio in the subject.

In some instances, the ANS is modulated in a manner sufficient to shiftor change parasympathetic activity and/or sympathetic activity from afirst state to a second state, where the second state is characterizedby an increase or decrease in the sympathetic activity/parasympatheticactivity ratio relative to the first state.

Accordingly, some embodiments of the subject invention includemodulating at least a portion of a subject's autonomic nervous system toincrease the sympathetic activity/parasympathetic activity ratio, i.e.,to increase sympathetic activity relative to parasympathetic activity(in other words to decrease parasympathetic activity relative tosympathetic activity) so as to treat a subject for a condition that canbe treated by such modulation (e.g., a condition caused byparasympathetic bias). Alternatively or in addition to stimulating atleast one sympathetic nerve fiber to increase activity, increasing thesympathetic activity/parasympathetic activity ratio may be achieved byinhibiting activity in the parasympathetic system.

Other embodiments of the subject invention include modulating asubject's autonomic nervous system to decrease the sympatheticactivity/parasympathetic activity ratio, i.e., to decrease sympatheticactivity relative to parasympathetic activity (in other words, toincrease parasympathetic activity relative to sympathetic activity) soas to treat a subject for a condition that can be treated by suchmodulation (e.g., a condition caused by sympathetic bias).

As will be described in greater detail below, while the ratio ofsympathetic function/parasympathetic function may be modulated accordingto embodiments of the subject invention to treat or improve a subjectfor a condition (e.g., aging associated conditions) the net result maybe a parasympathetic bias (i.e., a parasympathetic dominance), asympathetic bias (i.e., sympathetic dominance), or the activities of thesympathetic system and parasympathetic system may be substantially equal(i.e., neither is dominant).

By “bias”, is meant that the particular “biased” component of theautonomic nervous system has a higher activity level than the othercomponent. For example, a parasympathetic bias refers to a higher levelof parasympathetic activity than sympathetic activity, and vice versa,where such bias may be systemic or localized. As such, by “vagal bias”,is meant that that the particular biased component of the autonomicnervous system that has a higher activity level than the other componentis the vagus nerve or a portion of the autonomic nervous systemassociated with the vagus nerve. Vagal bias may be characterized by oneor more of vagal dominance, vagal hypersensitivity and/or sympatheticinsufficiency. The net result of the subject methods to treat acondition may be higher or greater sympathetic activity relative toparasympathetic activity in at least the area of the targeted autonomicsystem (i.e., that portion in need of modulation), or substantiallyequal activity levels of sympathetic activity and parasympatheticactivity.

As noted above, in certain embodiments activity in at least a portion ofthe autonomic nervous system is increased. For example, activity in atleast a portion of the ANS that is involved the sympathetic nervoussystem may be increased such that at least a portion of the sympatheticnervous system may be “up-regulated”. In other instances, any portion ofthe ANS that is involved in the parasympathetic system, e.g., one ormore nerve fibers, may be stimulated to increase parasympatheticactivity to provide the desired ratio of parasympathetic/sympatheticactivity. In other words, activity in at least a portion of theparasympathetic nervous system may be increased such that at least aportion of the parasympathetic nervous system may be “up-regulated”.

In certain embodiments, increasing activity in, or up-regulating, atleast a part of the sympathetic system may be desired in instanceswhere, prior to the application of autonomic nervous system-modulatingagent, parasympathetic activity is higher than desired, e.g., higherthan sympathetic activity (e.g., there exists a relative parasympatheticbias) and as such the subject methods may be employed to increasesympathetic activity to a level above or rather to a level that isgreater than parasympathetic activity or may be employed to modulate thedifferential between the parasympathetic-sympathetic systems such thatthe result of increasing sympathetic activity may be a sympathetic bias,parasympathetic bias or may be an equalization of the two systems (i.e.,the activities of the two systems are approximately equal—includingequal), but the difference between the parasympathetic-sympatheticsystems may be modulated, e.g., reduced or minimized or increased incertain embodiments. Accordingly, the subject methods may be employed toincrease sympathetic activity above that of parasympathetic activityand/or may be employed to modulate (increase or decrease) thedifferential between the two systems, but in certain embodiments may beemployed to decrease the parasympathetic activity/sympathetic activityratio.

In other embodiments, increasing activity in, or up-regulating, at leasta part of the parasympathetic system may be desired in instances where,prior to the application of autonomic nervous system-modulating agent,sympathetic activity is higher than desired, e.g., higher thanparasympathetic activity (e.g., there exists a relative sympatheticbias) and as such the subject methods may be employed to increaseparasympathetic activity to a level above or rather to a level that isgreater than sympathetic activity or may be employed to modulate thedifferential between the parasympathetic-sympathetic systems such thatthe result of increasing parasympathetic activity may be aparasympathetic bias, sympathetic bias or may be an equalization of thetwo systems (i.e., the activities of the two systems are approximatelyequal—including equal), but the difference between theparasympathetic-sympathetic systems may be modulated, e.g., reduced orminimized or increased in certain embodiments. Accordingly, the subjectmethods may be employed to increase parasympathetic activity above thatof sympathetic activity and/or may be employed to modulate (increase ordecrease) the differential between the two systems, but in certainembodiments may be employed to decrease the parasympatheticactivity/sympathetic activity ratio.

In certain embodiments, a parasympathetic bias may be the normal state,but the ratio of the two systems may be abnormal or otherwisecontributing to a condition. Increasing sympathetic bias may also bedesired in instances where, prior to the restoration of the normalfunction of a central nervous system endocrine gland, sympatheticactivity is higher than the parasympathetic activity, but thedifferential between the two needs to be modulated such as increasedfurther, e.g., the sympathetic activity is normal or above normal (i.e.,abnormally high) and/or the parasympathetic activity is normal or belownormal (i.e., abnormally low) or above normal (i.e., abnormally low).

For example, such instances may occur where a subject has normal orabove normal sympathetic function, but also has elevated parasympatheticfunction. Other instances may include below normal sympathetic function,but normal or elevated parasympathetic function, etc. It may also bedesirable to increase sympathetic function in instances where therespective activities of the two system are analogous or approximatelyequal, including equal, prior to increasing activity in the sympatheticsystem, but the level of one or both is abnormally high or abnormallylow. The above-described examples of instances where increasingsympathetic activity may be desired is exemplary only and is in no wayintended to limit the scope of the invention and other instances whereincreasing sympathetic activity may be desired will be apparent to thoseof skill in the art.

In other embodiments, a sympathetic bias may be the normal state, butthe ratio of the two systems may be abnormal or otherwise contributingto a condition. Increasing parasympathetic bias may also be desired ininstances where, prior to the restoration of the normal function of acentral nervous system endocrine gland, parasympathetic activity ishigher than the sympathetic activity, but the differential between thetwo needs to be modulated such as increased further, e.g., theparasympathetic activity is normal or above normal (i.e., abnormallyhigh) and/or the sympathetic activity is normal or below normal (i.e.,abnormally low) or above normal (i.e., abnormally low).

For example, such instances may occur where a subject has normal orabove normal parasympathetic function, but also has elevated sympatheticfunction. Other instances may include below normal parasympatheticfunction, but normal or elevated sympathetic function, etc. It may alsobe desirable to increase parasympathetic function in instances where therespective activities of the two system are analogous or approximatelyequal, including equal, prior to increasing activity in theparasympathetic system, but the level of one or both is abnormally highor abnormally low. The above-described examples of instances whereincreasing parasympathetic activity may be desired is exemplary only andis in no way intended to limit the scope of the invention and otherinstances where increasing sympathetic activity may be desired will beapparent to those of skill in the art.

As noted above, in certain embodiments, activity in at least a portionof the ANS may be inhibited to modulate at least a portion of theautonomic nervous system. Inhibiting or “down-regulating” activity in atleast a part of the autonomic nervous system, may be desired ininstances where, the sympathetic or parasympathetic activity is higherthan desired. For example, parasympathetic activity may be higher thanthe sympathetic activity (i.e., there exists a parasympathetic bias) orparasympathetic activity may be less than or approximately equal to,including equal, to sympathetic activity, and the subject methods may beemployed to modulate the differential between theparasympathetic-sympathetic systems such that the net result ofdecreasing sympathetic activity may be a sympathetic bias,parasympathetic bias or may be an equalization of the two systems (i.e.,the activities of the two systems are approximately equal—includingequal), but the difference between the parasympathetic-sympatheticsystems may be modulated, e.g., increased or reduced in certainembodiments. Accordingly, the subject methods may be employed todecrease parasympathetic activity below that of sympathetic activityand/or may be employed to modulate (decrease or increase) thedifferential between the two systems, where in certain embodiments maybe employed to decrease the ratio of parasympathetic activity tosympathetic activity.

For example, decreasing activity in at least a portion of theparasympathetic system may be employed where there is a normal or anabnormally low sympathetic function and/or abnormally highparasympathetic function. Such may also be desired in instances where,prior to decreasing parasympathetic function in, e.g., at least oneparasympathetic nerve fiber, sympathetic activity is higher than theparasympathetic activity, but the differential between the two needs tobe increased further. For example, such instances may occur where asubject has normal or above normal (i.e., abnormally high)parasympathetic function, but also has elevated sympathetic function(i.e., abnormally high), e.g., a relative bias towards sympatheticfunction may be present. Other instances include normal or below normal(i.e., abnormally low) parasympathetic activity and/or normal or abovenormal (i.e., abnormally high) sympathetic activity. The above-describedexamples of instances where decreasing parasympathetic activity may bedesired is exemplary only and is in no way intended to limit the scopeof the invention and other instances where decreasing parasympatheticactivity to provide an increase in the parasympatheticactivity/sympathetic activity ratio may be desired will be apparent tothose of skill in the art.

Decreasing activity in at least a portion of the sympathetic system maybe employed where there is a normal or an abnormally low parasympatheticfunction and/or abnormally high sympathetic function. Such may also bedesired in instances where, prior to decreasing sympathetic function in,e.g., at least one parasympathetic nerve fiber, parasympathetic activityis higher than the sympathetic activity, but the differential betweenthe two needs to be increased further. For example, such instances mayoccur where a subject has normal or above normal (i.e., abnormally high)sympathetic function, but also has elevated parasympathetic function(i.e., abnormally high), e.g., a relative bias towards parasympatheticfunction may be present. Other instances include normal or below normal(i.e., abnormally low) sympathetic activity and/or normal or abovenormal (i.e., abnormally high) parasympathetic activity. Theabove-described examples of instances where decreasing sympatheticactivity may be desired is exemplary only and is in no way intended tolimit the scope of the invention and other instances where decreasingsympathetic activity to provide an increase in the parasympatheticactivity/sympathetic activity ratio may be desired will be apparent tothose of skill in the art.

One way of inhibiting activity in at least a portion of the autonomicnervous system is by the application of a nerve block. Application of anerve block at least partially prevents nerve transmission across thelocation of the block. A nerve block can be administered to modulateautonomic function using all the methods and devices described hereinincluding pharmacological and/or electrical means. As noted above, incertain embodiments, activity in at least a portion of the autonomicnervous system may be increased and activity in at least a portion ofthe autonomic nervous system may be decreased. For example, in certainembodiments, activity in at least a portion of the sympathetic systemmay be increased and activity in at least a portion of theparasympathetic system may be inhibited, e.g., to decrease theparasympathetic activity/sympathetic activity ratio. In otherembodiments, activity in at least a portion of the parasympatheticsystem may be increased and activity in at least a portion of thesympathetic system may be inhibited, e.g., to decrease theparasympathetic activity/sympathetic activity ratio. As described above,any portion of the parasympathetic and/or sympathetic nervous systemsmay be modulated to increase activity and activity in any portion of theANS may be inhibited to provide the desired ratio of parasympatheticactivity to sympathetic activity. Such a protocol may be employed, e.g.,in instances where sympathetic function is normal or abnormally lowand/or parasympathetic function is normal or abnormally high, or whereparasympathetic function is normal or abnormally low and/or sympatheticfunction is normal or abnormally high, where normal is determined by thetypical or average autonomic nervous system functions for a healthysubject, e.g., a healthy human subject ranging in age from about 20years old to about 25 years old.

Embodiments wherein activity in at least a portion of the autonomicnervous system may be increased and activity in at least a portion ofthe autonomic nervous system may be decreased may be employed to alterthe dominance and/or may be employed to modulate the differentialbetween the two systems. For example, prior to modulating the autonomicsystem according to the subject invention, the activity in theparasympathetic system may be higher than activity in the sympatheticsystem and the subject methods may be employed to increase thesympathetic activity to a level that is greater than the parasympatheticactivity and/or may be employed to alter the differential or differencein activity levels of the two systems such as decreasing the differencein activity levels or increasing the difference in activity levels.

Increasing activity in at least a portion of the autonomic nervoussystem, e.g., increasing activity in at least a portion of theparasympathetic system, and decreasing activity in at least a portion ofthe autonomic nervous system, e.g., decreasing activity in at least aportion of the sympathetic system, may be performed simultaneously orsequentially such that at least a portion of the autonomic nervoussystem, e.g., at least a portion of the parasympathetic nervous system,may be pharmacologically and/or electrically modulated to increaseactivity therein prior or subsequent to inhibiting activity in at leasta portion of the autonomic nervous system e.g., at least a portion ofthe sympathetic nervous system, such as by electrical and/orpharmacological means.

Regardless of whether increasing activity in at least a portion of theautonomic nervous system, e.g., in at least a portion of theparasympathetic system, and decreasing activity in at least a portion ofthe autonomic nervous system, e.g., in at least a portion of thesympathetic system, is performed simultaneously or sequentially, theparameters for increasing activity in at least a portion of autonomicnervous system and decreasing activity in at least a portion of theautonomic nervous system may be analogous to that described above.

Modulation of the autonomic nervous system may be accomplished using anysuitable method, including employing electrical, thermal, vibrational,magnetic, acoustic, baropressure, optical, or other sources of energy tomodulate autonomic balance, where in representative embodimentsmodulation is achieved via pharmacological modulation and/or electricalenergy modulation in a manner that is effective to treat a subject for afood allergy syndrome condition.

Certain embodiments include pharmacologically or electricallystimulating a portion of the subject's nervous system in a manner thatcauses a modulation of at least a portion of a subject's autonomicnervous system, e.g., by increasing parasympathetic activity and/ordecreasing sympathetic activity or by increasing sympathetic activityand/or decreasing parasympathetic activity in at least a portion of thesubject's autonomic nervous system. In certain embodiments, modulationmay include increasing the sympathetic activity/parasympathetic activityratio in at least a portion of the subject's autonomic nervous system.In certain embodiments, a combination of electrical and pharmacologicalmay be employed.

Pharmacologic Modulation

In certain embodiments of the subject methods, the therapy comprises apharmacological modulation, which modulation may result in modulation ofthe ANS and/or some other system of the subject in manner effective tomodulate the dynamic measure of homeostatic capacity, as desired. By“pharmacologically modulation” is meant altering or changing one or moresystems of the subject by pharmacological means to provide a desiredchange, alteration or shift in system(s) function. In embodiments inwhich pharmacological agent is administered, any suitable protocol maybe used, where certain protocols include using an pharmacological agentadministering device to deliver a suitable amount of pharmacologicalagent to a subject. Methods and corresponding devices and systems forapplying at least one pharmacological agent to a subject and which maybe adapted for use in the subject invention are described, e.g., in U.S.Pat. Nos. 7,363,076; 7,149,574, 7,738,952; 7,899,527; 7,676,269;8,121,690; 8,569,277; 8,909,340 United States Published Application Nos.20050143378; 20100260669; 20110015188; 20100119482; 20110256097;20060206149; 20140065129; 20140369969 and U.S. patent application Ser.No. 14/737,248; the disclosures of which are herein incorporated byreference.

Any convenient pharmacological agent may be employed. Pro-sympatheticagents of interest include, but are not limited to: beta agonists, e.g.,dobutamine, metaproterenol, terbutaline, ritodrine, albuterol; alphaagonists, e.g., selective alpha 1-adrenergic blocking agents such asphenylephrine, metaraminol, methoxamine; prednisone and steroids, (e.g.,available under the brand names CORATN, DELTASONE, LIQUID PRED,MEDICORTEN, ORASONE, PANASOL-S, PREDNICEN-M, PREDNISONE INTENSOL);indirect agents that include norepinephrine, e.g., ephedrine,ampthetamines, phenylpropanolamines, cyclopentamines, tuaminoheptanes,naphazolines, tetrahydrozolines; epinephrine; norepinephrine;acetylcholine; sodium; calcium; angiotensin I; angiotensin II;angiotensin converting enzyme I (“ACE I”); angiotensin converting enzymeII (“ACE II”); aldosterone; potassium channel blockers and magnesiumchannel blockers, e.g., valproate (sodium valproate, valproic acid),lithium; cocaine; amphetamines; terbutaline; dopamine; doputamine;antidiuretic hormone (“ADH”) (also known as vasopressin); oxytocin(including PITOCINE); THC cannabinoids; and combinations thereof.

Pro-parasympathetic agents of interest include, but are not limited to:Beta Blockers, Aldosterone Antagonists; Angiotensin II ReceptorBlockers; Angiotensin Converting Enzyme Inhibitors; Statins;Triglyceride Lowering Agents; Insulin Sensitizers; InsulinSecretagogues; Insulin Analogs; Alpha-glucosidase Inhibitors; SGLT2Inhibitors; Immunomodulators, including agents that bind/react to CD4,gp39, B7, CD19, CD20, CD22, CD401, CD40, CD40L and CD23 antigens;Sympathomimetics; Cholinergics; Calcium Channel Blockers; Sodium ChannelBlockers; Glucocorticoid Receptor Blockers; Peripheral AdrenergicInhibitors; Blood Vessel Dilators; Central Adrenergic Agonists;Alpha-adrenergic Blockers; Combination Diuretics; Potassium-sparingDiuretics; Nitrate Pathway Modulators; Cyclic NucleotideMonophosphodiesterase (PDE) Inhibitors; Vasopressin Inhibitors; ReninInhibitors; Estrogen and Estrogen Analogues and Metabolites; VesicularMonoamine Transport (VMAT) Inhibitors; Progesterone Inhibitors;Testosterone Inhibitors; Gonadotropin-releasing Hormone Inhibitors;Dipeptidyl Peptidase IV inhibitors; Anticoagulants; Thrombolytics.

Pharmaceutical agents of interest also include biotherapeutic agents.Biotherapeutic agents include, but are not limited to: nucleic acidagents, polypeptide agents, complex biological preparations, e.g., bloodproducts and derivatives thereof, e.g., plasma, mitochondrialpreparations (e.g., for mitochondrial transfer); etc.

In some instances, the agent modulates the activity of the proteinfollowing expression, such that the agent is one that changes theactivity of the protein encoded by a target gene following expression ofthe protein from the target gene. In these instances, the agent is onethat may act directly with protein encoded by the target gene.

In yet other embodiments, the agent modulates expression of the RNAand/or protein from the gene, such that it changes the expression of theRNA or protein from the target gene in some manner. In these instances,the agent may change expression of the RNA or protein in a number ofdifferent ways. In certain embodiments, the agent is one that reduces,including inhibits, expression of a functional target protein.Inhibition of protein expression may be accomplished using anyconvenient means, including use of an agent that inhibits proteinexpression, such as, but not limited to: antisense agents, RNAi agents,agents that interfere with transcription factor binding to a promotersequence of the target gene, or inactivation of the target gene, e.g.,through recombinant techniques, etc.

For example, antisense molecules can be used to down-regulate expressionof a target gene in the cell. The anti-sense reagent may be antisenseoligodeoxynucleotides (ODN), particularly synthetic ODN having chemicalmodifications from native nucleic acids, or nucleic acid constructs thatexpress such anti-sense molecules as RNA. The antisense sequence iscomplementary to the mRNA of the targeted protein, and inhibitsexpression of the targeted protein. Antisense molecules inhibit geneexpression through various mechanisms, e.g., by reducing the amount ofmRNA available for translation, through activation of RNAse H, or sterichindrance. One or a combination of antisense molecules may beadministered, where a combination may include multiple differentsequences.

Antisense molecules may be produced by expression of all or a part ofthe target gene sequence in an appropriate vector, where thetranscriptional initiation is oriented such that an antisense strand isproduced as an RNA molecule. Alternatively, the antisense molecule is asynthetic oligonucleotide. Antisense oligonucleotides will generally beat least about 7, usually at least about 12, more usually at least about20 nucleotides in length, and not more than about 500, usually not morethan about 50, more usually not more than about 35 nucleotides inlength, where the length is governed by efficiency of inhibition,specificity, including absence of cross-reactivity, and the like. It hasbeen found that short oligonucleotides, of from 7 to 8 bases in length,can be strong and selective inhibitors of gene expression (see Wagner etal. (1996), Nature Biotechnol. 14:840-844).

A specific region or regions of the endogenous sense strand mRNAsequence is chosen to be complemented by the antisense sequence.Selection of a specific sequence for the oligonucleotide may use anempirical method, where several candidate sequences are assayed forinhibition of expression of the target gene in an in vitro or animalmodel. A combination of sequences may also be used, where severalregions of the mRNA sequence are selected for antisense complementation.

Antisense oligonucleotides may be chemically synthesized by methodsknown in the art (see Wagner et al. (1993), supra, and Milligan et al.,supra.) Oligonucleotides may be chemically modified from the nativephosphodiester structure, in order to increase their intracellularstability and binding affinity. A number of such modifications have beendescribed in the literature, which alter the chemistry of the backbone,sugars or heterocyclic bases.

Among useful changes in the backbone chemistry are phosphorothioates;phosphorodithioates, where both of the non-bridging oxygens aresubstituted with sulfur; phosphoroamidites; alkyl phosphotriesters andboranophosphates. Achiral phosphate derivatives include3′-O′-5′-S-phosphorothioate, 3′-S-5′-O-phosphorothioate,3′-CH₂-5′-O-phosphonate and 3′-NH-5′-O-phosphoroamidate. Peptide nucleicacids replace the entire ribose phosphodiester backbone with a peptidelinkage. Sugar modifications are also used to enhance stability andaffinity. The α-anomer of deoxyribose may be used, where the base isinverted with respect to the natural β-anomer. The 2′-OH of the ribosesugar may be altered to form 2′-O-methyl or 2′-O-allyl sugars, whichprovides resistance to degradation without comprising affinity.Modification of the heterocyclic bases must maintain proper basepairing. Some useful substitutions include deoxyuridine fordeoxythymidine; 5-methyl-2′-deoxycytidine and 5-bromo-2′-deoxycytidinefor deoxycytidine. 5-propynyl-2′-deoxyuridine and5-propynyl-2′-deoxycytidine have been shown to increase affinity andbiological activity when substituted for deoxythymidine anddeoxycytidine, respectively.

As an alternative to anti-sense inhibitors, catalytic nucleic acidcompounds, e.g. ribozymes, anti-sense conjugates, etc. may be used toinhibit gene expression. Ribozymes may be synthesized in vitro andadministered to the patient, or may be encoded on an expression vector,from which the ribozyme is synthesized in the targeted cell (forexample, see International patent application WO 9523225, and Beigelmanet al. (1995), Nucl. Acids Res. 23:4434-42). Examples ofoligonucleotides with catalytic activity are described in WO 9506764.Conjugates of anti-sense ODN with a metal complex, e.g.terpyridylCu(II), capable of mediating mRNA hydrolysis are described inBashkin et al. (1995), Appl. Biochem. Biotechnol. 54:43-56.

In addition, the transcription level of a target protein can beregulated by gene silencing using RNAi agents, e.g., double-strand RNA(Sharp (1999) Genes and Development 13: 139-141). RNAi, such asdouble-stranded RNA interference (dsRNAi) or small interfering RNA(siRNA), has been extensively documented in the nematode C. elegans(Fire, A., et al, Nature, 391, 806-811, 1998) and routinely used to“knock down” genes in various systems. RNAi agents may be dsRNA or atranscriptional template of the interfering ribonucleic acid which canbe used to produce dsRNA in a cell. In these embodiments, thetranscriptional template may be a DNA that encodes the interferingribonucleic acid. Methods and procedures associated with RNAi are alsodescribed in WO 03/010180 and WO 01/68836, all of which are incorporatedherein by reference. dsRNA can be prepared according to any of a numberof methods that are known in the art, including in vitro and in vivomethods, as well as by synthetic chemistry approaches. Examples of suchmethods include, but are not limited to, the methods described by Sadheret al. (Biochem. Int. 14:1015, 1987); by Bhattacharyya (Nature 343:484,1990); and by Livache, et al. (U.S. Pat. No. 5,795,715), each of whichis incorporated herein by reference in its entirety. Single-stranded RNAcan also be produced using a combination of enzymatic and organicsynthesis or by total organic synthesis. The use of synthetic chemicalmethods enables one to introduce desired modified nucleotides ornucleotide analogs into the dsRNA. dsRNA can also be prepared in vivoaccording to a number of established methods (see, e.g., Sambrook, etal. (1989) Molecular Cloning: A Laboratory Manual, 2nd ed.;Transcription and Translation (B. D. Hames, and S. J. Higgins, Eds.,1984); DNA Cloning, volumes I and II (D. N. Glover, Ed., 1985); andOligonucleotide Synthesis (M. J. Gait, Ed., 1984, each of which isincorporated herein by reference in its entirety). A number of optionscan be utilized to deliver the dsRNA into a cell or population of cellssuch as in a cell culture, tissue, organ or embryo. For instance, RNAcan be directly introduced intracellularly. Various physical methods aregenerally utilized in such instances, such as administration bymicroinjection (see, e.g., Zernicka-Goetz, et al. (1997) Development124:1133-1137; and Wianny, et al. (1998) Chromosoma 107: 430-439). Otheroptions for cellular delivery include permeabilizing the cell membraneand electroporation in the presence of the dsRNA, liposome-mediatedtransfection, or transfection using chemicals such as calcium phosphate.A number of established gene therapy techniques can also be utilized tointroduce the dsRNA into a cell. By introducing a viral construct withina viral particle, for instance, one can achieve efficient introductionof an expression construct into the cell and transcription of the RNAencoded by the construct.

In another embodiment, the target gene is inactivated so that it nolonger expresses a functional protein. By inactivated is meant that thegene, e.g., coding sequence and/or regulatory elements thereof, isgenetically modified so that it no longer expresses a functional targetprotein. The alteration or mutation may take a number of differentforms, e.g., through deletion of one or more nucleotide residues,through exchange of one or more nucleotide residues, and the like. Onemeans of making such alterations in the coding sequence is by homologousrecombination. Methods for generating targeted gene modificationsthrough homologous recombination are known in the art, including thosedescribed in: U.S. Pat. Nos. 6,074,853; 5,998,209; 5,998,144; 5,948,653;5,925,544; 5,830,698; 5,780,296; 5,776,744; 5,721,367; 5,614,396;5,612,205; the disclosures of which are herein incorporated byreference.

Also of interest in certain embodiments are dominant negative mutants oftarget proteins, where expression of such mutants in the cell result ina modulation, e.g., decrease, in target protein activity. Dominantnegative mutants are mutant proteins that exhibit dominant negativetarget protein activity. As used herein, the term “dominant negativeactivity” refers to the inhibition, negation, or diminution of certainparticular activities of a target protein, such as the apoptoticactivity of a target protein. Dominant negative mutations are readilygenerated for corresponding proteins. These may act by several differentmechanisms, including mutations in a substrate-binding domain; mutationsin a catalytic domain; mutations in a protein binding domain (e.g.multimer forming, effector, or activating protein binding domains);mutations in cellular localization domain, etc. A mutant polypeptide mayinteract with wild-type polypeptides (made from the other allele) andform a non-functional multimer. In certain embodiments, the mutantpolypeptide will be overproduced. Point mutations are made that havesuch an effect. In addition, fusion of different polypeptides of variouslengths to the terminus of a protein, or deletion of specific domainscan yield dominant negative mutants. General strategies are availablefor making dominant negative mutants (see for example, Herskowitz (1987)Nature 329:219, and the references cited above). Such techniques areused to create loss of function mutations, which are useful fordetermining protein function. Methods that are well known to thoseskilled in the art can be used to construct expression vectorscontaining coding sequences and appropriate transcriptional andtranslational control signals for increased expression of an exogenousgene introduced into a cell. These methods include, for example, invitro recombinant DNA techniques, synthetic techniques, and in vivogenetic recombination. Alternatively, RNA capable of encoding geneproduct sequences may be chemically synthesized using, for example,synthesizers. See, for example, the techniques described in“Oligonucleotide Synthesis”, 1984, Gait, M. J. ed., IRL Press, Oxford.

In yet other embodiments, the agent is an agent that modulates, e.g.,inhibits, target protein activity by binding to the target proteinand/or inhibiting binding of target protein to a second protein. Forexample, small molecules that bind to a target protein and inhibit itsactivity are of interest. Naturally occurring or synthetic smallmolecule compounds of interest include numerous chemical classes, suchas organic molecules, e.g., small organic compounds having a molecularweight of more than 50 and less than about 2,500 daltons. Candidateagents comprise functional groups for structural interaction withproteins, particularly hydrogen bonding, and typically include at leastan amine, carbonyl, hydroxyl or carboxyl group, preferably at least twoof the functional chemical groups. The candidate agents may includecyclical carbon or heterocyclic structures and/or aromatic orpolyaromatic structures substituted with one or more of the abovefunctional groups. Candidate agents are also found among biomoleculesincluding peptides, saccharides, fatty acids, steroids, purines,pyrimidines, derivatives, structural analogs or combinations thereof.Such molecules may be identified, among other ways, by employing thescreening protocols described below.

In yet other instances, the agent is an agent that increases theactivity of a protein, e.g., by increasing the amount of protein, e.g.,in a cell. For example, introduction of an expression vector encoding apolypeptide can be used to express the encoded product in cells lackingthe sequence, or to over-express the product. Various promoters can beused that are constitutive or subject to external regulation, where inthe latter situation, one can turn on or off the transcription of agene. These coding sequences may include full-length cDNA or genomicclones, fragments derived therefrom, or chimeras that combine anaturally occurring sequence with functional or structural domains ofother coding sequences. Alternatively, the introduced sequence mayencode an anti-sense sequence; be an anti-sense oligonucleotide; encodea dominant negative mutation, or dominant or constitutively activemutations of native sequences; altered regulatory sequences, etc.

A variety of methods can be used to construct expression vectorscontaining coding sequences and appropriate transcriptional andtranslational control signals for increased expression of an exogenousgene introduced into a cell. These methods include, for example, invitro recombinant DNA techniques, synthetic techniques, and in vivogenetic recombination. Alternatively, RNA capable of encoding geneproduct sequences may be chemically synthesized using, for example,synthesizers. See, for example, the techniques described in“Oligonucleotide Synthesis”, 1984, Gait, M. J. ed., IRL Press, Oxford.

A variety of host-expression vector systems may be utilized to express agenetic coding sequence. Expression constructs may contain promotersderived from the genome of mammalian cells, e.g., metallothioneinpromoter, elongation factor promoter, actin promoter, etc., frommammalian viruses, e.g., the adenovirus late promoter; the vacciniavirus 7.5K promoter, SV40 late promoter, cytomegalovirus, etc.

In mammalian host cells, a number of viral-based expression systems maybe utilized, e.g. retrovirus, lentivirus, adenovirus, herpesvirus, andthe like. In cases where an adenovirus is used as an expression vector,the coding sequence of interest may be ligated to an adenovirustranscription/translation control complex, e.g., the late promoter andtripartite leader sequence. This chimeric gene may then be inserted inthe adenovirus genome by in vitro or in vivo recombination. Insertion ina non-essential region of the viral genome (e.g., region E1 or E3) willresult in a recombinant virus that is viable and capable of expressingthe gene product in infected hosts (see Logan & Shenk, 1984, Proc. Natl.Acad. Sci. USA 81:3655-3659). Specific initiation signals may also berequired for efficient translation of inserted gene product codingsequences. These signals include the ATG initiation codon and adjacentsequences. Standard systems for generating adenoviral vectors forexpression on inserted sequences are available from commercial sources,for example the Adeno-X™ expression system from Clontech (Clontechniques(January 2000) p. 10-12).

In cases where an entire gene, including its own initiation codon andadjacent sequences, is inserted into the appropriate expression vector,no additional translational control signals may be needed. However, incases where only a portion of the gene coding sequence is inserted,exogenous translational control signals, including, perhaps, the ATGinitiation codon, must be provided. Furthermore, the initiation codonmust be in phase with the reading frame of the desired coding sequenceto ensure translation of the entire insert. These exogenoustranslational control signals and initiation codons can be of a varietyof origins, both natural and synthetic. The efficiency of expression maybe enhanced by the inclusion of appropriate transcription enhancerelements, transcription terminators, etc. (see Bittner et al., 1987,Methods in Enzymol. 153:516-544).

In representative embodiments, methods are used that achieve a highefficiency of transfection, and therefore circumvent the need for usingselectable markers. These may include adenovirus infection (see, forexample Wrighton, 1996, J. Exp. Med. 183: 1013; Soares, J. Immunol.,1998, 161: 4572; Spiecker, 2000, J. Immunol 164: 3316; and Weber, 1999,Blood 93: 3685); and lentivirus infection (for example, InternationalPatent Application WO000600; or WO9851810). Adenovirus-mediated genetransduction of endothelial cells has been reported with 100%efficiency. Retroviral vectors also can have a high efficiency ofinfection with endothelial cells, with reported infection efficienciesof 40-77%. Other vectors of interest include lentiviral vectors, forexamples, see Barry et al. (2000) Hum Gene Ther 11(2):323-32; and Wanget al. (2000) Gene Ther 7(3):196-200.

Viral vectors include retroviral vectors (e.g. derived from MoMLV, MSCV,SFFV, MPSV, SNV etc), lentiviral vectors (e.g. derived from HIV-1,HIV-2, SIV, BIV, FIV etc.), adeno-associated virus (AAV) vectors,adenoviral vectors (e.g. derived from Ad5 virus), SV40-based vectors,Herpes Simplex Virus (HSV)-based vectors etc. A vector construct mayinclude drug resistance genes (neo, dhfr, hprt, gpt, bleo, puro etc)enzymes (β-galactosidase, alkaline phosphatase etc) fluorescent genes(e.g. GFP, RFP, BFP, YFP) or surface markers (e.g. CD24, NGFr, Lyt-2etc).

The gene or protein may be introduced into tissues or host cells by anynumber of routes, including viral infection, microinjection, or fusionof vesicles. Jet injection may also be used for intra-muscularadministration, as described by Furth et al. (1992), Anal Biochem205:365-368. The DNA may be coated onto gold microparticles, anddelivered intradermally by a particle bombardment device, or “gene gun”as described in the literature (see, for example, Tang et al. (1992),Nature 356:152-154), where gold microprojectiles are coated with theDNA, then bombarded into skin cells.

Once the gene corresponding to a selected polynucleotide is identified,its expression can be regulated in the cell to which the gene is native.For example, an endogenous gene of a cell can be regulated by anexogenous regulatory sequence as disclosed in U.S. Pat. No. 5,641,670;the disclosure of which is herein incorporated by reference.

Also of interest in these embodiments is the administration of a targetprotein itself or active fragments, as well as mimetics, thereof.

In some instances, the active agent is configured to cross the bloodbrain barrier. For example, the active agent may be conjugated to amoiety that confers upon the active agent the ability to cross the bloodbrain barrier. Such a configuration allows for the targeting of theactive agent to tissues within the blood brain barrier. In someembodiments the subject moiety may be a peptide, e.g., acell-penetrating peptide. Suitable peptides that facilitate crossing ofthe blood brain barrier include, but are not limited to positivelycharged peptides with amphipathic characteristics, such as MAP, pAntp,Transportan, SBP, FBP, TAT₄₈₋₆₀, SynB1, SynB3 and the like.

In other embodiments, the subject moiety may be a polymer. Suitablepolymers that facilitate crossing of the blood brain barrier include,but are not limited to, surfactants such as polysorbate (e.g., Tween®20, 40, 60 and 80); poloxamers such as Pluronic® F 68; and the like. Insome embodiments, an active agent is conjugated to a polysorbate suchas, e.g., Tween® 80 (which is Polyoxyethylene-80-sorbitan monooleate),Tween® 40 (which is Polyoxyethylene sorbitan monopalmitate); Tween® 60(which is Polyoxyethylene sorbitan monostearate); Tween® 20 (which isPolyoxyethylene-20-sorbitan monolaurate); polyoxyethylene 20 sorbitanmonopalmitate; polyoxyethylene 20 sorbitan monostearate; polyoxyethylene20 sorbitan monooleate; etc. Also suitable for use are water solublepolymers, including, e.g.: polyether, for example, polyalkylene oxidessuch as polyethylene glycol (“PEG”), polyethylene oxide (“PEO”),polyethylene oxide-co-polypropylene oxide (“PPO”), co-polyethylene oxideblock or random copolymers, and polyvinyl alcohol (“PVA”); poly(vinylpyrrolidinone) (“PVP”); poly(amino acids); dextran, and proteins such asalbumin. Block co-polymers are suitable for use, e.g., a polyethyleneoxide-polypropylene oxide-polyethylene-oxide (PEO-PPO-PEO) triblockco-polymer (e.g., Pluronic® F68); and the like; see, e.g., U.S. Pat. No.6,923,986. Other methods for crossing the blood brain barrier arediscussed in various publications, including, e.g., Chen & Liu (2012)Advanced Drug Delivery Reviews 64:640-665.

The targeting moiety may be attached to the subject active agent via anyconvenient method. The targeting moiety may be attached to the activeagent via a single bond or a suitable linker, e.g., a PEG linker, apeptidic linker including one or more amino acids, or a saturatedhydrocarbon linker. A variety of linkers find use in the subjectmodified compounds.

In certain embodiments where targeting moieties or active agents aresmall molecule compounds, such compounds may contain, or be modified tocontain, an α-nucleophilic group that serves as a reactive partneruseful in conjugation to a compound disclosed herein. General methodsare known in the art for chemical synthetic schemes and conditionsuseful for synthesizing a compound of interest (see, e.g., Smith andMarch, March's Advanced Organic Chemistry: Reactions, Mechanisms, andStructure, Fifth Edition, Wiley-Interscience, 2001; or Vogel, A Textbookof Practical Organic Chemistry, Including Qualitative Organic Analysis,Fourth Edition, New York: Longman, 1978).

In certain embodiments where targeting moieties or active agents arepeptides, any convenient reagents and methods may be used to conjugatethe targeting moiety and subject active agent, for example, conjugationmethods as described in G. T. Hermanson, “Bioconjugate Techniques”Academic Press, 2nd Ed., 2008, solid phase peptide synthesis methods, orfusion protein expression methods. Reactive functional groups forconjugation of peptidic compounds, via an optional linker, include, butare not limited to: an azido group, an alkynyl group, a phosphine group,a cysteine residue, a C-terminal thioester, aryl azides, maleimides,carbodiimides, N-hydroxysuccinimide (NHS)-esters, hydrazides,PFP-esters, hydroxymethyl phosphines, psoralens, imidoesters, pyridyldisulfides, isocyanates, aminooxy-, aldehyde, keto, chloroacetyl,bromoacetyl, and vinyl sulfones.

Other variations of standard peptide coupling chemistry may be employed.Examples of peptide coupling reagents that can be used include, but notlimited to, DCC (dicyclohexylcarbodiimide), DIC(diisopropylcarbodiimide), di-p-toluoylcarbodiimide, BDP(1-benzotriazolediethylphosphate-1-cyclohexyl-3-(2-morpholinylethyl)carbodiimide), EDC(1-(3-dimethylaminopropyl-3-ethyl-carbodiimide hydrochloride), cyanuricfluoride, cyanuric chloride, TFFH (tetramethyl fluoroformamidiniumhexafluorophosphosphate), DPPA (diphenylphosphorazidate), BOP(benzotriazol-1-yloxytris(dimethylamino)phosphoniumhexafluorophosphate), HBTU(0-benzotriazol-1-yl-N,N,N′,N′-tetramethyluronium hexafluorophosphate),TBTU (O-benzotriazol-1-yl-N,N,N′,N′-tetramethyluroniumtetrafluoroborate), TSTU(O—(N-succinimidyl)-N,N,N′,N′-tetramethyluronium tetrafluoroborate),HATU(N-[dimethylamino)-1-H-1,2,3-triazolo[4,5,6]-pyridin-1-ylmethylene]-N-methylmethanaminiumhexafluorophosphate N-oxide), BOP-CI(bis(2-oxo-3-oxazolidinyl)phosphinic chloride), PyBOP((1-H-1,2,3-benzotriazol-1-yloxy)-tris(pyrrolidino)phosphoniumtetrafluorophopsphate), BrOP (bromotris(dimethylamino)phosphoniumhexafluorophosphate), DEPBT(3-(diethoxyphosphoryloxy)-1,2,3-benzotriazin-4(3H)-one) PyBrOP(bromotris(pyrrolidino)phosphonium hexafluorophosphate).

In certain embodiments where targeting moieties or active agents areoligonucleotides, any convenient reagents and methods may be used toconjugate the targeting moiety and subject active agent. For exampleconjugation methods described in P. Herdewijn, “OligonucleotideSynthesis” Humana Press, 2005, such as total stepwise solid-phasesynthesis methods, or methods utilizing incorporation of 2′-aldehydesfor use in ligation via hydrazine, oxime, or thiazolidine linkages. Inother cases, the oligonucleotide may be first conjugated, by methodswell known in the art, to a natural or synthetic amino acid such thatfunctional groups on the amino acid may be utilized for conjugation byany of the relevant peptide conjugation methods described herein.

In another embodiment where the targeting moiety is an antibody, theantibody may include a light chain polypeptide including a C-terminalamino acid extension, which extension includes a cysteine residue, wherethe agent is conjugated to the cysteine residue (directly or indirectly(e.g., via a linker)) of the C-terminal amino acid extension. In oneembodiment, conjugation method involves the preferential (or “biased”)conjugation of agent to the cysteine residue of the C-terminal aminoacid extension over a cysteine residue outside the C-terminal extension.In certain aspects, the conjugation includes conjugating a linker to asulfhydryl group of the cysteine residue, e.g., using maleimide reactionchemistry, haloacetyl reaction chemistry, pyridyl disulfide reactionchemistry, or any other suitable reaction chemistry as describedelsewhere herein. The methods of making the conjugate may furtherinclude reducing the sulfhydryl group of the cysteine residue prior tothe conjugating step, e.g., using a suitable reducing agent and reactionconditions as described above. An alternative embodiment of the presentdisclosure does not require a reduction step as the cysteine within thelight chain extension is already in a reduced state as a synthesisproduct.

In certain aspects, the agent is linked to the cysteine of theC-terminal extension using maleimide reaction chemistry. The maleimidegroup reacts specifically with sulfhydryl groups when the pH of thereaction mixture is between pH 6.5 and 7.5; the result is formation of astable thioether linkage. In more alkaline conditions (pH>8.5), primaryamines compete with thiols for reaction with maleimides, and alsoincreases the rate of hydrolysis of the maleimide group to anon-reactive maleamic acid. Maleimides do not react with tyrosines,histidines or methionines. Bioconjugation approaches that employmaleimide-based linkers are known and described in detail, e.g., inHermanson, G. T., Bioconjugate Techniques, 2nd ed. San Diego, Calif.Academic Press 2008; Aslam & Dent, Bioconjugation: Protein CouplingTechniques for the Biomedical Sciences, London Macmillan Reference Ltd1998; Kalia & Raines, Advances in Bioconjugation, Curr. Org. Chem.14(2):138-147; and elsewhere.

According to certain embodiments, the agent is linked to the cysteine ofthe C-terminal extension using haloacetyl reaction chemistry. In certainaspects, a haloacetyl crosslinker that includes an iodoacetyl or abromoacetyl group is employed. Haloacetyls react with sulfhydryl groupsat physiologic pH. The reaction of the iodoacetyl group proceeds bynucleophilic substitution of iodine with a sulfur atom from a sulfhydrylgroup, resulting in a stable thioether linkage.

In certain aspects, the agent is linked to the cysteine of theC-terminal extension using pyridyl disulfide reaction chemistry. Pyridyldisulfides react with sulfhydryl groups over a broad pH range (with pH 4to 5 being optimal) to form disulfide bonds. During the reaction, adisulfide exchange occurs between the molecule's —SH group and thereagent's 2-pyridyldithiol group. As a result, pyridine-2-thione isreleased and can be measured spectrophotometrically (Amax=343 nm) tomonitor the progress of the reaction.

To generate a reduced sulfhydryl in the cysteine of the C-terminal aminoacid extension to which the agent may be attached (e.g., via a linker),the sulfhydryl group of the cysteine may be contacted with a suitablereducing agent under conditions sufficient to reduce the sulfhydrylgroup. In certain aspects, the reducing agent is selected fromcysteamine hydrochloride, 2-mercaptoethanol, dithiothreitol (DTT),2-mercaptoethylamine, tris(2-carboxyl)phosphine (TCEP), cysteine HCl,N-ethylmaleimide, Nacystelyn, dornase alfa, thymosin 134, guaifenesinTCEP HCl, and any combination thereof. Reaction conditions for suchreducing agents are known in the art and may be optimized, e.g., topromote selectivity or “bias” the reduction of the sulfhydryl group ofthe cysteine(s) present in the C-terminal extension as opposed to thecysteine residues present in the parental antibody (e.g., the cysteineresidues that participate in disulfide bonding between CL and CH1 of thelight and heavy chains, and/or between the hinge regions of the heavychains). An alternative embodiment of the invention does not require areduction step as the cysteine within the light chain extension isalready in a reduced state as a synthesis product.

Preferential reduction of the cysteine(s) of the C-terminal amino acidextension over one or more cysteine residues outside the C-terminalamino acid extension (or exclusive reduction of the cysteine(s) of theC-terminal amino acid extension) may be achieved by selection ofsuitable reduction conditions. In certain aspects, suitable reductionconditions include suitable selection of one or more of the following: amild reducing agent and/or a reducing agent having a steric bulk thatconfers upon the reducing agent a preference for reducing a cysteine ofthe C-terminal amino acid extension; concentrations of the reducingagent and substrate; the temperature at which the reduction reaction iscarried out, the pH of the reduction reaction mixture; the buffer usedin the reduction reaction; and/or conditions under which the cellsexpressing the extended C-terminal light chain polypeptides are cultured(e.g., to obtain free thiol on the C-terminal extension and/or togenerate readily reduced intermolecular disulfides). The agentconjugated to the antibody may be any useful agent described elsewhereherein. In certain aspects where the agent is an antibody, the agent maybe conjugated to a targeting moiety by antibody conjugation methodsdescribed herein.

The ordinarily skilled artisan will appreciate that factors such as pHand steric hindrance (i.e., the accessibility of the amino acid residueto reaction with a reactive partner of interest) are of importance.Modifying reaction conditions to provide for optimal conjugationconditions is well within the skill of the ordinary artisan, and isroutine in the art.

In practicing methods according to embodiments of the invention, aneffective amount of the active agent is provided in the target cell orcells. In some instances, the effective amount of the modulatory agentis provided in the cell by contacting the cell with the modulatoryagent. Contact of the cell with the modulatory agent may occur using anyconvenient protocol. The protocol may provide for in vitro or in vivocontact of the modulatory agent with the target cell, depending on thelocation of the target cell. Contact may or may not include entry of theagent into the cell. For example, where the target cell is an isolatedcell and the modulatory agent is an agent that modulates expression oftarget protein, the modulatory agent may be introduced directly into thecell under cell culture conditions permissive of viability of the targetcell. Such techniques include, but are not necessarily limited to: viralinfection, transfection, conjugation, protoplast fusion,electroporation, particle gun technology, calcium phosphateprecipitation, direct microinjection, viral vector delivery, and thelike. The choice of method is generally dependent on the type of cellbeing contacted and the nature of the modulatory agent, and thecircumstances under which the transformation is taking place (e.g., invitro, ex vivo, or in vivo). A general discussion of these methods canbe found in Ausubel, et al, Short Protocols in Molecular Biology, 3rded., Wiley & Sons, 1995.

Alternatively, where the target cell or cells are part of amulticellular organism, the modulatory agent may be administered to theorganism or subject in a manner such that the agent is able to contactthe target cell(s), e.g., via an in vivo or ex vivo protocol. By “invivo,” it is meant in the target construct is administered to a livingbody of an animal. By “ex vivo” it is meant that cells or organs aremodified outside of the body. Such cells or organs are typicallyreturned to a living body.

In the subject methods, the active agent(s) may be administered to thetargeted cells using any convenient administration protocol capable ofresulting in the desired activity. Thus, the agent can be incorporatedinto a variety of formulations, e.g., pharmaceutically acceptablevehicles, for therapeutic administration. More particularly, the agentsof the present invention can be formulated into pharmaceuticalcompositions by combination with appropriate, pharmaceuticallyacceptable carriers or diluents, and may be formulated into preparationsin solid, semi-solid, liquid or gaseous forms, such as tablets,capsules, powders, granules, ointments (e.g., skin creams), solutions,suppositories, injections, inhalants and aerosols. As such,administration of the agents can be achieved in various ways, includingoral, buccal, rectal, parenteral, intraperitoneal, intradermal,transdermal, intracheal, etc., administration.

In pharmaceutical dosage forms, the agents may be administered in theform of their pharmaceutically acceptable salts, or they may also beused alone or in appropriate association, as well as in combination,with other pharmaceutically active compounds. The following methods andexcipients are merely exemplary and are in no way limiting.

For oral preparations, the agents can be used alone or in combinationwith appropriate additives to make tablets, powders, granules orcapsules, for example, with conventional additives, such as lactose,mannitol, corn starch or potato starch; with binders, such ascrystalline cellulose, cellulose derivatives, acacia, corn starch orgelatins; with disintegrators, such as corn starch, potato starch orsodium carboxymethylcellulose; with lubricants, such as talc ormagnesium stearate; and if desired, with diluents, buffering agents,moistening agents, preservatives and flavoring agents.

The agents can be formulated into preparations for injection bydissolving, suspending or emulsifying them in an aqueous or nonaqueoussolvent, such as vegetable or other similar oils, synthetic aliphaticacid glycerides, esters of higher aliphatic acids or propylene glycol;and if desired, with conventional additives such as solubilizers,isotonic agents, suspending agents, emulsifying agents, stabilizers andpreservatives.

The agents can be utilized in aerosol formulation to be administered viainhalation. The compounds of the present invention can be formulatedinto pressurized acceptable propellants such as dichlorodifluoromethane,propane, nitrogen and the like.

Furthermore, the agents can be made into suppositories by mixing with avariety of bases such as emulsifying bases or water-soluble bases. Thecompounds of the present invention can be administered rectally via asuppository. The suppository can include vehicles such as cocoa butter,carbowaxes and polyethylene glycols, which melt at body temperature, yetare solidified at room temperature.

Unit dosage forms for oral or rectal administration such as syrups,elixirs, and suspensions may be provided wherein each dosage unit, forexample, teaspoonful, tablespoonful, tablet or suppository, contains apredetermined amount of the composition containing one or moreinhibitors. Similarly, unit dosage forms for injection or intravenousadministration may comprise the inhibitor(s) in a composition as asolution in sterile water, normal saline or another pharmaceuticallyacceptable carrier.

The term “unit dosage form,” as used herein, refers to physicallydiscrete units suitable as unitary dosages for human and animalsubjects, each unit containing a predetermined quantity of compounds ofthe present invention calculated in an amount sufficient to produce thedesired effect in association with a pharmaceutically acceptablediluent, carrier or vehicle. The specifications for the novel unitdosage forms of the present invention depend on the particular compoundemployed and the effect to be achieved, and the pharmacodynamicsassociated with each compound in the host.

The pharmaceutically acceptable excipients, such as vehicles, adjuvants,carriers or diluents, are readily available to the public. Moreover,pharmaceutically acceptable auxiliary substances, such as pH adjustingand buffering agents, tonicity adjusting agents, stabilizers, wettingagents and the like, are readily available to the public.

Where the agent is a polypeptide, polynucleotide, analog or mimeticthereof, it may be introduced into tissues or host cells by any numberof routes, including viral infection, microinjection, or fusion ofvesicles. Jet injection may also be used for intramuscularadministration, as described by Furth et al. (1992), Anal Biochem205:365-368. The DNA may be coated onto gold microparticles, anddelivered intradermally by a particle bombardment device, or “gene gun”as described in the literature (see, for example, Tang et al. (1992),Nature 356:152-154), where gold microprojectiles are coated with theDNA, then bombarded into skin cells. For nucleic acid therapeuticagents, a number of different delivery vehicles find use, includingviral and non-viral vector systems, as are known in the art.

Those of skill in the art will readily appreciate that dose levels canvary as a function of the specific compound, the nature of the deliveryvehicle, and the like. Preferred dosages for a given compound arereadily determinable by those of skill in the art by a variety of means.

Electrical Modulation

In certain embodiments, to accomplish the desired modulation of thesubject dynamic measure of homeostatic capacity, electrical energy(electrical modulation) may be applied to at least a portion of asubject, where such electrical energy may be excitatory or inhibitoryand in certain embodiments may include both excitatory and inhibitorystimulation. By “electrically modulating” is meant altering or changingat least a portion of the subject by electrical means to provide achange, alteration or shift in at least one component or aspect of anelectrical system of the subject.

Any suitable area may be targeted for electrical modulation. Areas thatmay be targeted include, but are not limited to, pre- andpost-ganglionic nerve fibers, as well as ganglionic structures, efferentand afferent nerve fibers, synapses, etc., and combinations thereof incertain embodiments. In certain embodiments, activity in a given nervefiber may be electrically modulated in more than one area of the nervefiber. In certain embodiments, electrical energy is applied to modulatesynaptic efficiency. In certain embodiments, electrical energy isapplied using any of the devices described below.

A number of different methods and corresponding devices and systems forapplying electrical energy to a subject and which may be adapted for usein the subject invention are described, e.g., in U.S. Pat. Nos.: in U.S.Pat. Nos. 7,363,076; 7,149,574, 7,738,952; 7,899,527; 7,676,269;8,121,690; 8,569,277; 8,909,340 United States Published Application Nos.20050143378; 20100260669; 20110015188; 20100119482; 20110256097;20060206149; 20140065129; 20140369969 and U.S. patent application Ser.No. 14/737,248; the disclosures of which are herein incorporated byreference.

Paradoxical Modulation

In some instances, the methods include employing a paradoxical protocolin order to obtain a desired modulation in the dynamic measure ofhomeostatic capacity. In some of these embodiments, a counter-intuitivestimulus is applied to the subject in a manner effective to cause thesubject to mount a compensatory response effective to ultimatelymodulate the dynamic homeostatic capacity of the subject, as desired. Inpracticing methods according to such embodiments, a stimulus is appliedto the subject, where the stimulus is of a nature and magnitudesufficient to achieve the desired modulation. In certain embodiments,the applied stimulus is one of short duration, where by short durationis meant that the applied stimulus lasts for 1 week or less, e.g., 3days or less, e.g., 1 day or less, e.g., 12 hours or less, 5 hours orless, 1 hour or less, 30 min or less, 15 min or less, 5 min or less, 1min or less, 30 s or less, 1 s or less, where the duration of theapplied stimulus may be even shorter. In certain embodiments, theapplied stimulus is one of long duration, where by long duration ismeant that the applied stimulus lasts for 1 week or longer, e.g., 2weeks or longer, 1 month or longer, 2 months or longer, 3 months orlonger, or 6 months or longer, where the duration of the appliedstimulus may be even shorter. Where the stimulus is a pharmacologicalstimulus, the duration refers to the period in which the pharmacologicalagent from an administered dosage is active. Where the stimulus is anelectrical stimulus, the duration refers to the total of electricalapplications received by a subject over a given period, analogous to adose of a pharmacological agent.

Following administration, the stimulus is removed, e.g., bymetabolization of the pharmacological agent or cessation of applicationof electrical energy, and the subject is permitted to mount acompensatory response. In this following period, no additional stimulusis administered to the subject. The duration of this period betweenstimulus application, which may be referred to as a “holiday” period,may vary, but in representative embodiments is 1 second or longer, suchas 30 seconds or longer, e.g., 1 minute or longer, 5 minutes or longer,10 minutes or longer, 15 minutes or longer, 30 minutes or longer, 1 houror longer, 6 hours or longer, 12 hours or longer, 1 day or longer, suchas 2 days or longer, including 5 days or longer, 10 days or longer,e.g., 15 days or longer. As such, embodiments of the methods includenon-chronic (i.e., non-continuous) application of the stimulus, e.g.,non-chronic administration of a pharmacologic agent.

In certain embodiments, the methods include close monitoring orsupervision of the subject during and/or after application of thestimulus. This monitoring may be completely automated, or at least inpart performed manually, e.g., by a health care professional. Forexample, a health care professional can closely watch the subjectfollowing application of the stimulus as well as during the holidayperiod following stimulus application, and based on this monitoringdetermine when a next stimulus should be applied. Monitoring alsoassures that the symptom enhancement is not so severe as to beultimately damaging to the subject at an unacceptable level. Certainaspects of the monitoring may be automated. For example, followingadministration, the subject may enter one or more physiologicalparameters into an automated system, which uses the input parameters toautomatically determine whether the subject is staying within apredetermined set of physiological parameters, or whether interventionis necessary. In certain embodiments, the automated monitoring systemmay also be integrated with a stimulus application device, such that thesystem, based on monitored parameters, determines when next toadminister a stimulus, the duration of the next stimulus, etc. As such,the method may be characterized as applying a first stimulus to thesubject and monitoring the subject for a response thereto. Followingthis first step, the method further includes applying at least a secondstimulus to the subject, wherein the second stimulus is determined basedon the monitored response to the first stimulus.

In certain embodiments, stimulus to the subject is done in an“irregularly irregular” manner. As such, duration of the stimulusapplication events, as well as duration of holiday periods between suchevents, varies randomly over the entire course of a treatment, or atleast a portion thereof. In addition, the variation does not follow anypattern, but instead is random.

In practicing the subject methods, the applied stimulus may vary, wherein certain embodiments the stimulus may be a pharmacological stimulusand/or an electrical stimulus. As such, in certain embodiments, thestimulus is a pharmacological stimulus. In other embodiments, thestimulus is an electrical stimulus. In yet other embodiments, thestimulus is a combination of pharmacological and electrical stimuli.Accordingly, in certain embodiments, the enhancing is by administering apharmacological agent to the subject. In yet other embodiments, theenhancing is by electrical stimulation, e.g., by employing an implantedelectrical energy application device.

Further details regarding paradoxical therapies that may be employed inembodiments of the methods include those described in U.S. Pat. Nos.8,691,877 and 8,571,650, the disclosures of which are hereinincorporated by reference.

Pulsatile Therapy

In some instances, the methods include employing a pulsatile protocol inorder to obtain a desired modulation in the dynamic measure ofhomeostatic capacity. In some of these embodiments, a stimulus isapplied in a pulsatile manner to the subject effective to cause thedesired modulation in homeostatic capacity. Pulsatile protocols may beemployed to enhance homeostatic capacity and aspects thereof, e.g.,dynamic range, robustness, etc. In pulsatile stimulation protocols,intermittent stressors may be employed, e.g., in the form of iterativestress and rest and/or variation (irregularity or regularity) andintermittency of stressor, e.g., in order to enhance homeostaticcapacity. A dynamic range of stressors may be employed to increase thedynamic range of homeostatic capacity and/or to strengthen homeostaticcapacity.

In practicing methods according to such embodiments, a pulsatilestimulus is applied to the subject, where the pulsatile stimulus is of anature and magnitude sufficient to achieve the desired modulation. Incertain embodiments, the applied pulsatile stimulus is one of shortduration, where by short duration is meant that the applied stimuluslasts for 1 week or less, e.g., 3 days or less, e.g., 1 day or less,e.g., 12 hours or less, 5 hours or less, 1 hour or less, 30 min or less,15 min or less, 5 min or less, 1 min or less, 30 s or less, 1 s or less,where the duration of the applied stimulus may be even shorter. Incertain embodiments, the applied pulsatile stimulus is one of longduration, where by long duration is meant that the applied stimuluslasts for 1 week or longer, e.g., 2 weeks or longer, 1 month or longer,2 months or longer, 3 months or longer, or 6 months or longer, where theduration of the applied stimulus may be even shorter. Where thepulsatile stimulus is a pharmacological stimulus, the duration refers tothe period in which the pharmacological agent from an administereddosage is active. Where the pulsatile stimulus is an electricalstimulus, the duration refers to the total of electrical applicationsreceived by a subject over a given period, analogous to a dose of apharmacological agent.

Following administration of a given stimulation in a pulsatile stimulusprotocol, there is a non-stimulation period. The duration of thisnon-stimulation period between stimuli application, which may bereferred to as a “holiday” period, may vary, but in certain embodimentsis 1 second or longer, such as 30 seconds or longer, e.g., 1 minute orlonger, 5 minutes or longer, 10 minutes or longer, 15 minutes or longer,30 minutes or longer, 1 hour or longer, 6 hours or longer, 12 hours orlonger, 1 day or longer, such as 2 days or longer, including 5 days orlonger, 10 days or longer, e.g., 15 days or longer.

In certain embodiments, pulsatile stimulus to the subject is done in an“irregularly irregular” manner. As such, duration of the stimulusapplication events, as well as duration of holiday periods between suchevents, varies randomly over the entire course of a treatment, or atleast a portion thereof. In addition, the variation does not follow anypattern, but instead is random.

In practicing the subject methods, the applied pulsatile stimulus mayvary, where in certain embodiments the pulsatile stimulus may be apharmacological stimulus and/or an electrical stimulus. As such, incertain embodiments, the stimulus is a pharmacological stimulus. Inother embodiments, the stimulus is an electrical stimulus. In yet otherembodiments, the stimulus is a combination of pharmacological andelectrical stimuli. Accordingly, in certain embodiments, the enhancingis by administering a pharmacological agent to the subject. In yet otherembodiments, the enhancing is by electrical stimulation, e.g., byemploying an implanted electrical energy application device.

Behavioral Therapy

In some embodiments, the therapy that is administered to the subject isa behavioral therapy. By “behavioral therapy” is meant at protocol orregimen that results in a change in the behavior, i.e., the way that thesubject acts, in a manner sufficient to modulate the dynamic measure ofhomeostatic capacity and treat the subject for the target condition.Behavioral therapies that may be employed may vary, where examples ofsuch therapies include, but are not limited to: exercise regimens (e.g.,cardiovascular, weight lifting, stretching, yoga); resting/sleepingregimens (e.g., meditation); physical therapies; psychologicaltherapies, e.g., counseling for enhancement of emotions/mood; substanceabuse therapies, e.g., smoking cessation therapies, alcohol abstinencetherapies; drugs of abuse abstinence therapies, etc. Behavioraltherapies may vary in terms of application, where examples include butare not limited to those that are administered via professional and/orconsumer devices/services, e.g., mobile apps, videos, computers, etc.

Dietary Therapy

In some embodiments, the therapy that is administered to the subject isa dietary therapy. By “dietary therapy” is meant at protocol or regimenthat results in a change in the nutritional and/or chemical intake ofthe subject, e.g., the types of foods/liquids that the subject ingestsor otherwise introduces into the body, in a manner sufficient tomodulate the dynamic measure of homeostatic capacity and treat thesubject for the target condition. Dietary therapies that may be employedmay vary, where examples of such therapies include, but are not limitedto: low carbohydrate diets, low fat diets, low calorie diets, vegetariandiets, organic diets, etc.; nutritional supplement regimens, e.g.,vitamin regimens; etc.

Environment Therapy

In some embodiments, the therapy that is administered to the subject isan environmental therapy. By “environmental therapy” is meant atprotocol or regimen that results in a change in the contextualenvironment of the subject, e.g., the perceived surroundings of thesubject, in a manner sufficient to modulate the dynamic measure ofhomeostatic capacity and treat the subject for the target condition.Environmental therapies that may be employed may vary, where examples ofsuch therapies include, but are not limited to: changes in day/nightduration; changes in geographic locations, e.g., to obtain a desiredtemperature and/or elevation, etc.

Surgical Therapy

In some embodiments, the therapy that is administered to the subject isa surgical therapy. By “surgical therapy” is meant a manual or operativeprocedure on a living subject. Surgical procedures may vary widely, andmay or may not be minimally invasive, as is known in the art.

Modulation of Dynamic Measure of Homeostatic Capacity

As summarized above, the therapy is administered to the subject (e.g.,by a health practitioner and/or the subject itself, depending the natureof the particular therapy) in a manner sufficient to modulate thesubject's dynamic measure of homeostatic capacity to more closelyapproximate a target dynamic measure of homeostatic capacity and treatthe subject for the condition. In some embodiments, the methods resultin an enhancement or an increase in the dynamic measure of thehomeostatic capacity of the subject. The magnitude of theenhancement/increase may vary, where in some instances the magnitude is2-fold or greater, such as 5-fold or greater, e.g., 10-fold or greater.

In some embodiments, the methods may result in at least partiallyrestoring the dynamic measure of homeostatic capacity of the subject. By“at least partially restoring the homeostatic capacity of the subject”is meant that the homeostatic capacity of the subject is restored to benormal, e.g., in those embodiments were normal is the target dynamicmeasure. By “normal” is meant the dynamic measure of homeostaticcapacity of a healthy subject of a particular age. In certainembodiments, the healthy subject is a healthy human at an age afterpuberty, e.g., 18 year old, 19 year old, 20 year old, 21 year old, 22year old, 23 year old, 24 year old, 25 year old, 26 year old, 27 yearold, 28 year old, 29 year old, 30 year old, 31 year old, 32 year old, 33year old 34 year old, 35 year old, 36 year old, 37 year old, 38 yearold, 39 year old, 40 year old, 41 year old, 42 year old, 43 year old, 44year old, 45 year old, 46 year old, 47 year old, 48 year old, 49 yearold or 50 year old. In some instances, the normal function with respectto homeostatic capacity is that of a healthy human 25 year old. In someinstances, the dynamic measure is enhanced to a target dynamic measurethat is greater than that observed in a normal subject, e.g., a supernormal value. In these instances, the magnitude by which the targetdynamic measure may exceed the normal measure may vary, such as by2-fold or greater, e.g., 5-fold or greater, including 10-fold orgreater.

As indicated above, the therapies are applied such that the dynamicmeasure of homeostatic capacity more closely approximates a targetdynamic measure, e.g., the normal measure or super normal measure, suchas described above. By “approximates” is meant, in some instances, thatthe dynamic measure of homeostatic capacity is changed by the therapy tobe 50% or more, e.g., 75% or more of the target function, such as 80% ormore of the target dynamic measure, including 90% or more of the targetfunction, e.g., 95% or more of the target function, including 99% ormore of the target dynamic measure.

Therapeutic methods as described herein may further include, followingapplication of therapy, assessing dynamic homeostatic capacity todetermine with the measure approximates the target measure, as desired.In such embodiments, the subject's dynamic measure of homeostaticcapacity may be made using any convenient protocol, such as thatdescribed above.

Devices and Systems

A number of different devices and systems may be employed in accordancewith the subject invention. Devices and systems that may be adapted orconfigured for use in the subject invention include devices and systemsfor obtaining dynamic biometric data from a subject and optionallyfurther processing the obtained data in some, e.g., e.g., in making ahomeostatic capacity evaluation of the subject based on the obtaineddynamic biometric data, in making a dynamic diagnosis based on theobtained dynamic biometric data, etc. In some instances, the device maybe configured to also output a therapeutic treatment regimenrecommendation based on the homeostatic capacity evaluation and/orprovide such a therapeutic treatment to the subject.

Devices of interest may include one or more functional modules, whichmay be distributed among two or more distinct hardware units orintegrated into a single hardware unit, e.g., as described in greaterdetail below. In some instances, the devices include a dynamic biometricdata obtainment module, a homeostatic capacity evaluation module, and ahomeostatic capacity evaluation output module. The dynamic biometricobtainment module is adapted to obtain dynamic biometric data, e.g., bybeing in operational communication with one or more biometric parametersensors and or an input configured to receive dynamic biometric datafrom a source of such data, and transmit the obtained biometric data tothe process unit module. The homeostatic capacity evaluation module isadapted to retrieve the dynamic biometric data from the dynamicbiometric data obtainment module and make a homeostatic capacityevaluation therefrom. As such, the module is configured to produce ahomeostatic capacity evaluation from the received or input dynamicbiometric data. In some instances, the systems further include atherapeutic treatment regimen module, which is configured to identify asuitable therapeutic regimen based on the homeostatic capacityevaluation. The output module is adapted to provide the homeostaticcapacity evaluation (and in some instances a therapeutic treatmentregimen) to a user, e.g., the subject or interested stakeholder. In someinstances, the output module is configured to display the homeostaticcapacity evaluation to a user, e.g., via graphical user interface (GUI).In one embodiment, a visual display can be used for displaying thehomeostatic capacity evaluation. Other outputs may also be employed,e.g., printouts, messages (e.g., text messages or emails) sent toanother display device, to a storage location for later viewing (e.g.,the cloud), etc.

One embodiment of a device for evaluating a subject's homeostaticcapacity is configured as follows. A dynamic biometric obtainment moduleis configured to obtain subject's dynamic biometric data. This biometricdata from the subject may then be input into a homeostatic capacityevaluation module, along with biometric data from a database, whichcontains data made up from individuals of a variety of different agesand health of known homeostatic capacities. The homeostatic capacityevaluation module evaluates the subject's homeostatic capacity based onthe biometric data from the subject and from the database using aclassification rule derived from a machine learning algorithm, which maybe any convenient algorithm, such as but not limited to: Fisher's lineardiscriminant, logistic regression, naïve Bayes classifier, quadraticclassifiers, k-nearest neighbor, decision trees, neural networks, andsupport vector machine. The homeostatic capacity evaluation module maythen output the subject's predicted homeostatic capacity in auser-readable format via a homeostatic capacity evaluation outputmodule.

An example of a device according to an embodiment of the invention asdescribed above is illustrated in the flow chart of FIG. 1. Dynamicbiometric obtainment module 100 is adapted to obtain subject's dynamicbiometric data 110. This biometric data 110 from the subject is theninput into the homeostatic capacity evaluation module 140, along withbiometric data 130 from a database 120. The database 120 contains datamade up from individuals of a variety of different ages and health ofknown homeostatic capacities. The homeostatic capacity evaluation module140 evaluates the subject's homeostatic capacity based on the biometricdata from the subject 110 and from the database 130 using aclassification rule derived from a machine learning algorithm, which maybe any convenient algorithm, such as but not limited to: Fisher's lineardiscriminant, logistic regression, naïve Bayes classifier, quadraticclassifiers, k-nearest neighbor, decision trees, neural networks, andsupport vector machine. The homeostatic capacity evaluation outputmodule 150 then provides the homeostatic capacity evaluation to theuser.

FIG. 2 illustrates aspects of the device of FIG. 1 in greater detail,including implementation of a machine learning algorithm in order toclassify subjects according to their homeostatic capacities. Biometricdata comprising a training set 210 is obtained from a database 200,which contains classified or labeled, training examples with biometricvalues. In other words, database 200 has biometric data from individualsof known homeostatic capacities. The training set biometric data 210 isinput into a machine learning algorithm 250 of a homeostatic capacityevaluation module 240. A user 220 may define the type ofclassification/machine learning algorithm 230 to be used. The machinelearning algorithm 250 is optimized using one of a variety ofstatistical means known in the art, such as cross-validation.Alternatively (not shown), the user may define a plurality of machinelearning algorithms, or the computer may define a plurality of machinelearning algorithms, for which optimization methods will be performedand the best (most accurate) will be used. Once the machine learningalgorithm 250 is optimized, a classification rule 260 is established.Dynamic biometric obtainment module 270 is adapted to obtain subject'sdynamic biometric data 280. This biometric data 280 from the subject isthen input into the classification rule 260 of the homeostatic capacityevaluation module 240. The subject's homeostatic capacity is evaluatedusing the classification rule 260. The predicted homeostatic capacityclassification/evaluation is provided to the user by the homeostaticcapacity evaluation output module 290.

As would be recognized by one of skilled in the art, many differentsoftware, firmware, hardware options and data structures can be employedin devices of the invention, e.g., as described above. In someinstances, a general-purpose computer can be configured to a functionalarrangement for the methods and programs disclosed herein. The hardwarearchitecture of such a computer is well known by a person skilled in theart, and can comprise hardware components including one or moreprocessors (CPU), a random-access memory (RAM), a read-only memory(ROM), an internal or external data storage medium (e.g., hard diskdrive). A computer system can also comprise one or more graphic boardsfor processing and outputting graphical information to display means.The above components can be suitably interconnected via a bus inside thecomputer. The computer can further comprise suitable interfaces forcommunicating with general-purpose external components such as amonitor, keyboard, mouse, network, etc. In some embodiments, thecomputer can be capable of parallel processing or can be part of anetwork configured for parallel or distributive computing to increasethe processing power for the present methods and programs. In someembodiments, the program code read out from the storage medium can bewritten into a memory provided in an expanded board inserted in thecomputer, or an expanded unit connected to the computer, and a CPU orthe like provided in the expanded board or expanded unit can actuallyperform a part or all of the operations according to the instructions ofthe program code, so as to accomplish the functions described below. Inother embodiments, the method can be performed using a cloud computingsystem. In these embodiments, the datafiles and the programming can beexported to a cloud computer, which runs the program, and returns anoutput to the user.

The memory of a computer system can be any device that can storeinformation for retrieval by a processor, and can include magnetic oroptical devices, or solid-state memory devices (such as volatile ornon-volatile RAM). A memory or memory unit can have more than onephysical memory device of the same or different types (for example, amemory can have multiple memory devices such as multiple drives, cards,or multiple solid state memory devices or some combination of the same).With respect to computer readable media, “permanent memory” refers tomemory that is permanent. Permanent memory is not erased by terminationof the electrical supply to a computer or processor. Computer hard-driveROM (i.e., ROM not used as virtual memory), CD-ROM, floppy disk and DVDare all examples of permanent memory. Random Access Memory (RAM) is anexample of non-permanent (i.e., volatile) memory. A file in permanentmemory can be editable and re-writable. Operation of the computer iscontrolled primarily by operating system, which is executed by a centralprocessing unit. The operating system can be stored in a system memory.In some embodiments, the operating system includes a file system. Inaddition to the operating system, one possible implementation of thesystem memory includes a variety programming files and data files forimplementing the method described above.

Where desired, the devices may include one or more sensors, e.g.,configured to obtain biometric data, e.g., as described above. Incertain aspects, a sensor includes one or more, such as a set of two ormore, such as two or three, electrodes that provide for sensing. Forexample, the electrodes may be configured to generate electrocardiogramdata. Alternatively, physiological sensors distinct from electrodes maybe included in the device. For example, a temperature sensor, such as athermistor, CMOS temperature sensor, resistive temperature devices(RTDs), may be employed to obtain precise measurements of temperature.An additional physiological sensor may include an LED and a photodiodecombined into a pulse oximeter, which may be employed to measure bloodoxygenation, which would also give information about pulse pressure. Thedevice may also include analyte detection sensors. For example, specificchemical sensors may be incorporated into the devices to detect thepresence of various agents, e.g., alcohol, glucose, BNP (B-typeNatriuretic peptide, which is associated with cardiac disease), etc.Sensors of interest include those configured to detect the presence of achemical analyte in a biological fluid sample, where analytes ofinterest include, but are not limited to: blood sugar (glucose),cholesterol, bilirubin, creatine, various metabolic enzymes, hemoglobin,heparin, hematocrit, vitamin K or other clotting factors, uric acid,carcinoembryonic antigen or other tumor antigens, various reproductivehormones such as those associated with ovulation or pregnancy, drugs ofabuse and/or metabolites thereof; blood alcohol concentration, etc. Incertain aspects, substances or properties for which the receiver isconfigured to detect include lactate (important for athletes), oxygen,pH, alcohol, tobacco metabolites, and illegal drugs (important for bothmedical diagnosis and law enforcement). Where the devices includes ananalyte detecting sensing element, this sensing element can beconfigured in the receiver in a number of different ways. For example, asensor that includes a selectively permeable membrane which is permeableto the agent one wants to detect may be provided, where there is anisolated cell behind the membrane and the agent passes through themembrane. Changes in the properties, such as electrical properties, ofthe cell, are then measured. In certain aspects, a small reservoir onthe side of the devices with a membrane across it is employed, andelectrical circuitry behind it is measured. Also of interest are ChemFETsensors, which are based on the binding of analyte to the sensor causinga change in the conductivity. In certain aspects, a material whoseelectrical properties (or other properties) are changed when thematerial, e.g., protein analyte, binds to it are employed. Blood alcoholconcentration may be determined any number of ways, including but notlimited to: sensors that analyze fluid samples, such as perspiration,optical spectroscopic sensors, etc.

Of interest are receivers that include at least an electrocardiography(ECG) sensor module. An ECG sensor module is a module which isconfigured to obtain ECG data and, if desired, additionally perform oneor more of processing the data in some way, storing the data andretransmitting the data. The ECG data may be employed by the receiver toderive a number of different metrics, including but not limited to:R-wave, heart rate, heart rate variability, respiration rate, etc. Wherethe device includes one or more physiological sensing functionalities,the device may further include sensing modules that are configured toobtain and process data from these sensing functionalities. For example,where the device includes an ECG sensing functionality, the device mayinclude an appropriate functional module (for example in the form ofprogramming) that can handle and process the raw data from thesesensors.

In use, dynamic biometric data information is input into the system, anda user receives a homeostatic capacity evaluation from the system, e.g.,as described above. In certain embodiments, instructions in accordancewith the method (e.g., in the form of a mobile app or other type ofstructure) described herein can be coded onto a computer-readable mediumin the form of “programming”, where the term “computer readable medium”as used herein refers to any storage or transmission medium (includingnon-transitory version so such) that participates in providinginstructions and/or data to a computer for execution and/or processing.Programming may take the form of any convenient algorithms. In someinstances, programming may include statistical analysis. Any of avariety of statistical methods known in the art and described herein,can be used, where statistical methods of interest include, for example,discriminant analysis, classification analysis, cluster analysis,analysis of variance (ANOVA), regression analysis, regression trees,decision trees, nearest neighbor algorithms, principal components,factor analysis, ensemble learning, AdaBoost, ALOPEX, analogicalmodeling, cascading classifiers, case-based reasoning, classifierchains, co-training, information fuzzy networks, logic learning machine,perceptron, multidimensional scaling and other methods of dimensionalityreduction, likelihood models, hypothesis testing, kernel densityestimation and other smoothing techniques, cross-validation and othermethods to guard against overfitting of the data, the bootstrap andother statistical resampling techniques, artificial intelligence,including artificial neural networks, machine learning, data mining, andboosting algorithms, and Bayesian analysis, etc.

Examples of storage media include a floppy disk, hard disk, opticaldisk, magneto-optical disk, CD-ROM, CD-ft magnetic tape, non-volatilememory card, ROM, DVD-ROM, Blue-ray disk, solid state disk, and networkattached storage (NAS), whether or not such devices are internal orexternal to the computer. A file containing information can be “stored”on computer readable medium, where “storing” means recording informationsuch that it is accessible and retrievable at a later date by acomputer. The computer-implemented method described herein can beexecuted using programming that can be written in one or more of anynumber of computer programming languages. Such languages include, forexample, Java (Sun Microsystems, Inc., Santa Clara, Calif.), VisualBasic (Microsoft Corp., Redmond, Wash.), and C++ (AT&T Corp.,Bedminster, N.J.), as well as any many others.

As mentioned above, the functional modules may be performed by a varietyof different hardware, firmware and software configurations. In someinstances, the functional modules will be distributed among a system oftwo or more distinct devices, e.g., mobile devices, remote devices (suchas cloud server devices), laboratory instrument devices, etc., which maybe in communication with each other, e.g., via wired or wirelesscommunication. In other instances, the distinct functional modules willbe integrated into a single device. Where the distinct functionalmodules are integrated into a single device, the device may have avariety of configurations. For example, the device may be a laboratorydevice, which may or may not be configured to a bench top device. In yetother instances, the device may be a handheld device, e.g., a smartphoneor tablet type device. In yet other instances, the device may be awearable device, such as a watch type device, a wearable patch typedevice, etc.

Embodiments of the invention may employ virtual reality components andvirtual reality mediated protocols. Virtual reality (both display andinput devices) or other such simulators may be employed. Virtual realityand similar simulators may be employed to collect data (e.g., throughthe use of various biometric sensors that work in conjunction withvirtual reality systems, such as measuring heart rate, eye movements andblink rates, measuring electrical impulses on the head (e.g., ear, neck,etc.), brain waves, etc.). Virtual reality and similar simulators may beemployed to apply various stimuli (e.g., create apsychological/physiological condition such as fear of heights, etc., andthe attendant increases in blood pressure, psychological distress,etc.). Virtual reality and similar simulators may be employed to intherapeutic embodiments, for example, to induce physiological, chemical,electrical, behavioral, and or psychological change—e.g., to overcomephobias, to reduce blood pressure/treat hypertension (e.g., viaparadoxically elevating pressure), treat depression, improve mood andwell-being, improve system balance or ability to restore balance, etc.In such instances, any convenient virtual reality input devices orsimilar simulators, incorporating one or more of the senses, may beemployed, e.g., to improve homeostatic capacity at all system levels.Suitable virtual reality systems include consumer use at home, at retaillocations (as a service), or medical grade, e.g., that are configured tobe used in clinic settings or at home. Where desired, the virtualreality systems may be connected to other devices, such an exercisemachine with various biometric monitors to collect data, apply stimuliand be used for therapy. These could be used for any number of medicalindications, performance enhancement for athletes, general consumerwellness use, etc.

In addition, the present invention contemplates the storage and accessto information present thereon, e.g., concerning homeostatic capacityevaluation, treatment regimen, therapeutic administration, etc., wheresuch access may be public or via an appropriate secured and privatesetting, e.g., wherein HIPAA standards are followed, such that thesystem may be HIPAA compliant.

All publications and patent applications mentioned in this specificationare herein incorporated by reference to the same extent as if eachindividual publication or patent application was specifically andindividually indicated to be incorporated by reference.

The invention now being fully described, it will be apparent to one ofskill in the art that many changes and modifications can be made theretowithout departing from the spirit and scope of the appended claims.

What is claimed is:
 1. A method of treating a subject for a condition,the method comprising: applying a stimulus to and withdrawing thestimulus from the subject; obtaining a panel of dynamic measures ofhomeostatic capacity for the subject during the application of thestimulus and after withdrawing the stimulus by obtaining dynamicbiometric data from different biological systems of the subject;evaluating the panel of dynamic measures of homeostatic capacity of thesubject based on age-based normative values for said measures ofhomeostatic capacity; and administering a therapy to the subject in amanner sufficient to modulate the panel of the subject's differentdynamic measures of homeostatic capacity to more closely approximatetarget dynamic measures of homeostatic capacity and treat the subjectfor the condition.
 2. The method according to claim 1, wherein thedynamic biometric data comprises biometric data obtained over a periodof time.
 3. The method according to claim 2, wherein the biometric datais continuously obtained over the period of time.
 4. The methodaccording to claim 1, wherein the dynamic biometric data is obtained byevaluating a biometric parameter for a rate of change over a period oftime.
 5. The method according to claim 1, wherein the method comprisesphysically monitoring the subject to obtain the dynamic biometric data.6. The method according to claim 1, wherein the method comprisesanalyzing a sample from the subject to obtain the dynamic biometricdata.
 7. The method according to claim 1, wherein the therapy comprisesa traditional medical therapy.
 8. The method according to claim 1,wherein the condition is a disease condition.
 9. The method according toclaim 8, wherein the disease condition is an aging-associated diseasecondition.
 10. The method according to claim 1, wherein the condition isaging.
 11. The method according to claim 1, wherein the subject is amammal.
 12. The method according to claim 11, wherein the subject is aprimate.
 13. The method according to claim 12, wherein the subject is ahuman.
 14. The method according to claim 11, wherein the subject is alaboratory research animal.
 15. The method according to claim 1, whereinthe panel of dynamic measures of homeostatic capacity comprises two ormore of baroreceptor sensitivity, heart rate variability, flow-mediatedvasodilation, postprandial glucose and triglyceride tolerance tests,gait balance, pupillary light reflex, and recovery time after cardiacstress.
 16. The method according to claim 1, wherein the stimulus is aphysical stimulus comprising a change in orientation of the subject,exercise, a change in temperature experienced by the subject, or acombination thereof.